NUCLEIC ACID-BINDING CHIPS FOR DETECTING NITROGEN DEFICIENCIES AS PART OF BIOPROCESS CONTROL

Abstract

nucleic acid-binding chips for monitoring bioprocesses, specifically for detecting nitrogen deficiencies. Said chips carry probes that are sensitive to at least three of the following 50 genes: kdgR, citA, htrA, ycn1, yppF, trpB, ggt, alsR, glnA, nrgA, yciC, yvtA, nrgB, ycnJ, glnR, yvlA, yncE, yvlB, trpF, ydfS, trpD, ycnK, trpB, trpC, nasD, ycdH, nasC, nasB, trpE, pckA, nasF, yrkC, and tnrA or the homolgs to SEQ ID NO: 91, 41, 53, 19, 55, 47, 21, 17, 9, 85, 45, 49, 95, 63, 15, 93, or 81 at a maximum of 80 different probes that are specific of nitrogen metabolism. The invention also relates to the use of corresponding gene probes, especially on the aforementioned chips, to corresponding methods and possible uses.

Description

[0001]The present invention relates to nucleic acid-binding chips for monitoring bioprocesses, specifically for detecting nitrogen deficiencies, as well as the use of corresponding gene probes, especially on the aforementioned chips, to corresponding methods and possible uses, which depend on such probes and chips.

[0002]In the industrial utilization of biological processes, one is faced with the very fundamental problem of monitoring their progress, of achieving the desired result, of conserving resources and/or of achieving an optimal result in a given time. Biological processes are understood to mean, for example, the culture of microorganisms on an agar plate or in a shaker culture, particularly however, their fermentation or the production of raw materials by the fermentation of microorganisms. There is extensive prior art for this in regard to both single cell eukaryotes, such as yeasts or streptomycetes, as well as to Gram-negative or Gram-positive bacteria.

[0003]Processes of this kind are monitored firstly by observing the properties and requirements of the concerned organisms, which change in the course of the process, these changes being reflected, for example, in the optical density and viscosity of the medium, in absorbed or released gases, in pH changes or in changing nutrient requirements. The measurement of enzymic activities via suitable assays, for example the detection of activities of interest in the culture supernatant, may also be included here.

[0004]Secondly, various techniques have been developed in recent years in order to monitor the metabolic processes of the organisms in question at the level of gene expression. A common method for this is the use of genes for readily detectable proteins as indicators of the activity of the promoters of the actual genes of interest (promoter analysis, gene expression analysis). For this, appropriate apparatuses ("(bio)sensors") have also been developed.

[0005]Other techniques are concerned with the detection of the proteins of interest, or of the mRNA coding for these proteins. These techniques include (1.) proteome analysis, i.e. observing the change in provision of the cells in question with proteins, which analysis is usually carried out by way of two-dimensional gel electrophoresis of cell lysates, (2.) analysis of the mRNA formed (transcriptome) by way of a "genomic DNA array" generated in an analogous manner, and (3.) chip technology.

[0006]The last technique is in a comparatively early stage of development. Whereas the two methods mentioned first are ultimately based on quantitative isolation procedures and time-consuming analyses of the macromolecules in question, chip technology is based on the principle of attaching probes for proteins or for nucleic acids on physically readable carriers (chips), which probes respond immediately to the presence of the proteins or nucleic acids in question. Compared to the two former technologies, chips of this kind promise to provide a real-time analysis of the concerned process (At-line analysis). Another advantage is the need for comparatively small amounts of samples.

[0007]The principle of chip-based measurements is presented, for example, diagrammatically in FIG. 2 of the article "Real-time electrochemical monitoring: toward green analytical chemistry" by J. Wang (Acc. Chem. Res.; ISSN 0001-4842; Rec. Sep. 12, 2001, pp. A-F). According to this, the sample to be analyzed is contacted with a biorecognition layer which may be, for example, an enzyme, an antibody, a receptor or DNA; the signal received thereby is emitted as voltage or electric potential via a transducer, for example an amperometric or potentiometric electrode, through an amplifier (amplification/processing). The study in question also mentions optical systems, compared to which the electronically analyzable systems were regarded by the author as being superior in regard to miniaturizability and other advantages.

[0008]Owing to the present invention, the protein-specific chips need not be considered. mRNA-recognizing chips are usually doped with complementary DNA molecules or DNA-analogs. Their manufacture and utilization for very detailed problems, such as for example the differentiation of point mutations, is described for example in the application WO 95/11995 A1. The DNA chip analyses include those with PCR amplification of the target sequence and those without amplification. There are also those with optical evaluation of the signals attributable to the recognition and those with electrical evaluation.

[0009]The optical detection methods partly require a mechanism for amplifying the signals. For this purpose, for example, fluorophores, acridinium esters or indirect detection via secondary binding events, for example via biotin, avidin/streptavidin or digoxigenin, have been described. In the last case, optical detection makes use of digoxigenin-specific antibodies that are labeled with an enzyme. Here, the enzyme activity is detected either colorimetrically or by way of luminescence. According to Westin et al. (2000), Nature Biotechnol., 18, pp. 199-204, hybridization may be coupled with a PCR on the DNA chip in order to be able to carry out the entire detection reaction on one chip ("lab-on-a-chip concept").

[0010]Other studies have described the development of DNA chips that miniaturize the principle of capillary electrophoresis for DNA sequencing or separation (Woolley and Mathies (1994), Proc. Natl. Acad. Sci., 91, pp. 11348-11352; Liu et al. (2000), Proc. Natl. Acad. Sci. 97, pp. 5369-5374).

[0011]Electrically readable DNA chips have already been presented in principle in some publications (Hoheisel (1999), DECHEMA Jahresbericht 1999, pp. 8-11; Hintsche et al. (1997), EXS, 80, pp. 267-283). Wright et al. (2000; Anal. Biochem., 282, pp. 70-79) utilized an ion channel sensor (ICS) for DNA detection, as has been described for the first time by Cornell et al. (1997: Nature, 387, pp. 580-583). This is a process in which the conductivity of molecular ion channels is detected by means of a binding reaction. The sensor is essentially an impedance element. According to Cheng et al. (1998; Nat. Biotechnol, 16, pp. 541-546), it is possible to utilize electrical pulses for amplifying the hybridization reaction on optical DNA chips. Fritsche et al. (2002; Laborwelt II) proposed an electrical chip system which employs metallic nanoparticles bound to oligonucleotides, for example. In this system, "metallic amplification" during the hybridization reaction causes a decrease in the electrical resistance at the electrode, and can then be measured as a signal.

[0012]Another approach is based on an electrical detection principle which uses DNA probes that, due to labeling with a suitable enzyme (e.g. alkaline phosphatase), after hybridization result in an electrically active substrate that can then be detected via a redox reaction at the electrode (Hintsche et al. (1997), EXS, 80, pp. 267-283).

[0013]If in regard to the fundamental configuration and the evaluation system it is decided to use a particular nucleic acid-recognition chip type, the more specific problem arises as to which gene activities are to be observed. For technical reasons, the number of genes that can be analyzed simultaneously using one nucleic acid chip is limited. Thus, in regard to the number of probes that can be applied to the chip, optically readable chips are currently superior to those, which can be evaluated electrically. The limits of the latter chips are determined by the miniaturizability of the electronic measuring units.

[0014]Thus the biological problem arises, as to which gene activities suitably depict the concerned process. This also includes monitoring product formation, if, for example, said product is produced by fermentation. At the same time, control genes may also be included which indicate if the process develops in a direction, which is not intended. In the course of this monitoring, for reasons of practicability, the number of different genes observed should not be too high.

[0015]Biotechnological processes involving Gram-positive bacteria are of particular industrial interest. These bacteria, particularly owing to their secretion capability, are used for the industrial production of valuable substances. Among said bacteria, those of the genus Bacillus and among these in turn the species B. subtilis, B. amyloliquefaciens, B. agaradherens, B. licheniformis, B. lentus and B. globigii are currently economically the most important.

[0016]The studies presented below, for example, are concerned with the simultaneous observation of the activity of a plurality of genes in bacteria (multiparametric recording). The article "Monitoring of genes that respond to process-related stress in large-scale bioprocesses" by Schweder et al. (1999), Biotech. Bioeng., 65, pp. 151-159, describes the alteration in mRNA levels of various stress factor-inducible genes, namely clpB, dnaK (induced during heat shock), uspA (glucose deficiency), proU (osmotic stress), pfl and frd (O₂ deficiency) and ackA (glucose surplus) in the course of fermentation of E. coli and during the subsequent concentration phase. They were recorded by means of a PCR-based method carried out in a conventional manner. In this connection, different rates of expression were detected already at various sites in the reactor, as were responses to altered conditions, which took place in a matter of seconds.

[0017]Another fermentation of E. coli is described in the study "Monitoring of genes that respond to overproduction of an insoluble recombinant protein in Escherichia coli glucose-limited fed-batch fermentations" by Jurgen et al. (2000), Biotech. Bioeng., 70, pp. 217-224. Here, expression of the genes lon, dnaK, ibpB, htrA, ppiB, groEL, tig, s6, 19 and dps is observed partly at the mRNA level, partly at the protein level, partly at both levels. The investigation was carried out by means of 2D PAGE and the DNA array technique. In view of the results, it was suggested to monitor recombinant bioprocesses such as heterologous protein preparation via (directly) process-concerned proteins and reporter genes such as ibpB.

[0018]The study "Genomic analysis of high-cell-density recombinant Escherichia coli fermentation and "cell conditioning" for improved recombinant protein yield" by R. T. Gill et al. (2001; Biotech. Bioeng., 72, pp. 85-95) is concerned with another observation of the course of a fermentation in which a recombinant protein is expressed by E. coli. This study describes increased expression of the stress genes degP, uvrB, alpA, mltB, recA, ftsH, ibpA, aceA and groEL under the conditions mentioned with high cell density, compared to low cell density. Said genes were grouped among each other into certain clusters, according to the strength of the reaction. This was determined via an approach based on RT-PCR and DNA microarray, which was supplemented by dot blot analysis and which was applied to samples from two points in time of the fermentation, that is at the beginning, at low cell density, and towards the end, at high cell density. From this, cell conditioning approaches were developed in order to reduce the stress response of the cells.

[0019]Fundamental differences in the expression patterns of Gram-positive organisms compared to those of Gram-negative bacteria are reviewed by the study "Proteome and transcriptome based analysis of Bacillus subtilis cells overproducing an insoluble heterologous protein" by Jurgen et al. (2001), Appl. Microbiol. Biotechnol, 55, pp. 326-332. The study describes the expression inter alia of the genes dnaK, groEL, grpE, clpP, clpC, clpX, rpsB and rplJ in B. subtilis, as can be determined via the DNA macroarray technique and via two-dimensional polyacrylamide gel electrophoresis. According to this, the genes for purine synthesis and pyrimidine synthesis and those of particular ribosomal proteins are expressed in Gram-positive bacteria used for overexpression more strongly than was to be expected from the findings with Gram-negative bacteria. Another difference relates to the proteases Lon and Clp.

[0020]Since that time, several of these genes or even nucleic acid-binding chips with several of these genes have been disclosed, or at least the possibility of their manufacture revealed, in a plurality of publications. Thus, for example, the two patent applications DE 10136987 A1 and DE 10108841 A1 disclose in each case a gene from Corynebacterium glutamicum, namely clpC and citB, respectively. Both genes are described as being relevant for the amino acid metabolism, this being the reason for an intended, commercially interesting utilization of said genes, which comprises inactivating or at least attenuating said genes in order to optimize the fermentative production of amino acids by this microorganism. According to these applications, further possible applications may consist of providing probes for the gene products in question on nucleic acid-binding chips.

[0021]On the other hand, more genomic data of various organisms are increasingly published, which contain such an abundance of sequence data that a representative selection therefrom seems desirable. Thus the patent application WO 02/055655 A2 discloses more than 1800 DNA sequences, which have been determined by completely sequencing the genome of the microorganism Methylococcus capsulatus.

[0022]Since then, for example, the complete genome of the gram-positive Bacillus licheniformis has also been sequenced. It is described in the publication "The Complete Genome Sequence of Bacillus licheniformis DSM13, an Organism with Great Industrial Potential" (2004) by B. Veith et al. in J. Mol. Microbiol. Biotechnol., 7 (4), pp. 204 to 211, and in addition is available under the entry AE017333 (Bases 1 bis 4.222.645) in the databank GeneBank (National Center for Biotechnology Information NCBI, National Institutes of Health, Bethesda, Md., USA; http://www.ncbi.nlm.nih.gov; at the date of Feb. 12, 2004).

[0023]By using the technique of optically analysable chips it is now even possible to manufacture nucleic acid-binding chips that reveal an almost complete genome or the associated transcriptome (genomic DNA chips).

[0024]With the application WO 2004/027092 A2, a representative cross section with a manageable number of genes is made available so as to identify various physiological states that an observed microorganism can pass through in the course of cultivation. They include, for example, states of starvation relating to various nutrients or stress situations such as, for example, heat shock or cold shock, shearing stress, oxidative stress or oxygen limitation. It concerns the following genes. acoA, ahpC, ahpF, citB, clpC, clpP, codY, cspA, cspB, des, dnaK, eno, glnR, groEL, groL, gsiB, ibpA, ibpB, katA, katE, lctP, ldh, opuAB, phoA, phoD, pstS, purC, purN, pyrB, pyrP, sigB, tnrA, trxA and ydjF. The associated DNA sequences of B. subtilis, E. coli and/or B. licheniformis are also found in this application. This also permitted the corresponding nucleic acid-binding chips to be manufactured, which when monitoring a bioprocess based on microorganisms, particularly for Gram-positive or Gram-negative bacteria, indicate changes in the metabolic activities characterizing said process.

[0025]Nucleic acid-binding chips that are based on this selection of genes furnish a certain, although overall rather only a rough outline of the concerned metabolic situations. Generally, they are not capable of specifically illuminating an individual partial problem; indeed a single positive signal can result from various situations or can also be a false positive, which is why--particularly so in such an ambiguous situation--it is sensible to separately analyze a chosen metabolic aspect. Indeed, for electrically readable nucleic acid-binding chips that have the advantage of a real-time analysis, the number of simultaneously available sites is limited, such that it is not simply additional gene probes that can be added for the analysis of specific metabolic situations.

[0026]Nucleic acid-binding chips for monitoring bioprocesses that are specifically focussed on the detection of phosphate deficiency situations or glucose deficiency situations are disclosed in the not previously published applications DE 102004061664.7 and DE 102005022145.9. They each carry a limited number of probes to indicate these specific stress situations.

[0027]A metabolic situation that can be critical for microorganisms and consequently limiting for a corresponding bioprocess, is that of nitrogen deficiency. Nitrogen is an essential element for nucleic acids and for amino acids and consequently for proteins, which themselves represent the major functional carriers and nutrients of biological organisms. Nitrogen deficiency therefore stands for a deficient provision of nutrient and can rapidly lead to the dying off of the cells or can at least prevent the achievement of higher cell densities. A nitrogen deficiency also affects the rate of product formation, especially during the fermentative production of proteins, such as for example industrial enzymes. Consequently, there exists a particular need in this regard to carry out a chip-based real-time analysis and because of the thereby rapidly obtained results, to be able to selectively and therefore more purposefully engage in the ongoing bioprocess. In this way a loss of product, which would result from an unrecognised or tardily recognized nitrogen shortage, is prevented.

[0028]Accordingly, the object was to identify genes that in organisms, especially microorganisms, can be associated as unambiguously as possible with the stress signal of nitrogen deficiency. The aim was to develop probes for these genes in order to be able to employ said probes to monitor corresponding bioprocesses.

[0029]Consequently, it should be possible to occupy nucleic acid-binding chips with gene probes for single or a plurality of these genes and thereby arrive at nucleic acid-binding chips that dependably notify the signal "nitrogen deficiency" in the course of a monitored bioprocess (nitrogen deficiency sensors). This object posed itself especially for those nucleic acid-binding chips, whose number of occupiable sites, due to their type of construction, is comparatively small, especially the electrically readable chips. Having said that, these chips have the advantages of a rapid readability and therefore permit an At-line analysis. This guarantees an if needed early intervention to optimise the bioprocess in question in regard to the nutrient supply.

[0030]Such a DNA-binding chip should be applicable in a plurality of comparable processes and be customized with comparatively minor variations for specific application possibilities. It should preferably be directed to bioprocesses based on Bacillus species, particularly B. subtilis, B. amyloliquefaciens, B. lentus, B. globigii, and quite particularly on B. Licheniformis. Among bioprocesses, special emphasis was on fermentations, in particular the industrial manufacture of products, quite particularly of overexpressed proteins.

[0031]A nitrogen deficiency sensor of this kind should also make possible corresponding processes for measuring the physiological state of the concerned cells and also corresponding possible uses for monitoring the concerned biological processes.

[0032]To achieve this object, a large number of genes from the biologically important bacterium B. licheniformis was investigated (Example 1) in regard to their activatability by converting the concerned culture into a state of nitrogen deficiency. Here it was surprisingly determined that by far not all genes involved in nitrogen metabolism deliver a well defined signal in this regard. In addition, likewise surprisingly, there was observed an activation of such genes that had not been brought into contact with the nitrogen metabolism, for example the gene for a putative malate synthesis, or those that code for proteins with a still unknown function. Furthermore, those genes were discounted from among these, which clearly respond to more than only this one stress signal. The genes identified in this way should now be considered, if required independently from their previously known function, according to the invention as nitrogen metabolic genes. This is supported by Example 2 of the present invention. Inductions of very different strengths were observed in this case. According to the teaching of the present invention, genes should be all the more suitable as indicators, the stronger this response. According to the invention, those genes are therefore selected as the nitrogen deficiency indicators, which provide a clear signal that is significantly above a defined threshold value. The more the result lies above this limit, the more they are to be regarded as a part of the solution of the inventive problem, and makes clear the corresponding grading of the preferred inventive aspects (see below).

[0033]In Table 1 are shown all 210 genes from Bacillus licheniformis DSM13 that were investigated in Example 1, whose induction under nitrogen deficiency was observed, wherein a factor of at least three was regarded as being significant. From these, all 49 genes shown in Table 2 are those whose induction by nitrogen deficiency amounted to at least the factor 8 at any of the measured times and for which it could be concluded (from experiments not shown here) that they were comparatively specific for this signal. In Table 3 they are again listed with respect to the strength of their observed maximum induction. Their DNA and amino acid sequences are listed in the sequence listing of the present application, wherein the odd numbers stand for DNA sequences and the following even numbers stand respectively for the derived amino acid sequences. The concerned SEQ ID numbers in Tables 2 and 3 also refer to these sequences. The following genes are concerned in the order of decreasing strength of the induction initiated by nitrogen deficiency (see Tables 2 and 3): [0034]gene (homolog to SEQ ID NO. 81, 82) coding for a hypothetical protein of unknown function; [0035]yrkC (unknown function--similar to proteins of unknown function; SEQ ID NO. 83, 84); [0036]nasF (uroporphyrine-III C-methyltransferase; SEQ ID NO. 39, 40); [0037]gene (homolog to SEQ ID NO. 93, 94) coding for a putative protein (putative nitrogen regulation protein P-II); [0038]pckA (phosphoenol pyruvate-carboxykinase; SEQ ID NO. 43, 44); [0039]trpE (anthranilate-synthase; SEQ ID NO. 67, 68); [0040]nasB (electron-transfer-subunit of the assimilatory nitrate reductase; SEQ ID NO. 33, 34); [0041]gene (homolog to SEQ ID NO. 15, 16) coding for a hypothetical protein (close homolog to the aldehyde-dehydrogenase DhaS); [0042]nasC (catalytic subunit of the assimilatory nitrate reductase; SEQ ID NO. 35, 36); [0043]ycdH (unknown function--similar to the binding protein of the ABC transporter; SEQ ID NO. 65, 66); [0044]gene (homolog to SEQ ID NO. 63, 64) coding for a putative protein (putative ammonium transporter); [0045]nasD (sub unit of the assimilatory nitrite reductase; SEQ ID NO. 37, 38); [0046]gene (homolog to SEQ ID NO. 95, 96) coding for a hypothetical protein of unknown function; [0047]gene (homolog to SEQ ID NO. 49, 50) coding for a putative protein (putative Na(+) bonded D-alanine-glycine permease); [0048]gene (homolog to SEQ ID NO. 45, 46) coding for a putative protein (putative malate synthase); [0049]gene (homolog to SEQ ID NO. 85, 86) coding for a putative protein (putative glycosyl hydrolase/lysozyme); [0050]gene (homolog to SEQ ID NO. 9, 10) coding for a putative protein (putative serine protease); [0051]gene (homolog to SEQ ID NO. 17, 18) coding for a putative protein (putative transcription regulator); [0052]trpC (indole-3-glycerine phosphate synthase; SEQ ID NO. 71, 72); [0053]trpB (beta-sub unit of tryptophan synthase; SEQ ID NO. 75, 76); [0054]ycnK (unknown function--similar to the transcription regulator of the DeoR family; SEQ ID NO. 31, 32); [0055]gene (homolog to SEQ ID NO. 21, 22) coding for a putative protein (putative phage capsid protein); [0056]trpD (anthranilate-phosphoribosyl transferase; SEQ ID NO. 69, 70); [0057]ydfS (unknown function--similar to proteins of unknown function; SEQ ID NO. 79, 80); [0058]trpF (phosphoribosyl-anthranilate isomerase; SEQ ID NO. 73, 74); [0059]gene (homolog to SEQ ID NO. 47, 48) coding for a putative protein (putative hydrolase); [0060]yvlB (unknown function--similar to proteins of unknown function; SEQ ID NO. 89, 90); [0061]yncE (unknown function; SEQ ID NO. 87, 88); [0062]yvlA (unknown function; SEQ ID NO. 97, 98); [0063]glnR (transcription repressor of the glutamine synthetase gene; SEQ ID NO. 11, 12); [0064]ycnJ (unknown function--similar to the copper export protein; SEQ ID NO. 29, 30); [0065]gene (homolog to SEQ ID NO. 55, 56) coding for a putative protein (ATP binding protein of a putative ABC transporter); [0066]gene (homolog to SEQ ID NO. 19, 20) coding for a conserved hypothetical protein of unknown function; [0067]nrgB (nitrogen regulating PII-similar protein; SEQ ID NO. 3, 4); [0068]yvtA (unknown function--similar to the HtrA-similar serine protease; SEQ ID NO. 23, 24); [0069]yciC (unknown function--similar to proteins of unknown function; SEQ ID NO. 57, 58); [0070]nrgA (ammonium transporter; SEQ ID NO. 5, 6); [0071]glnA (glutamine synthetase; SEQ ID NO. 13, 14); [0072]alsR (transcription regulator of the alpha-acetolactate operon; SEQ ID NO. 61, 62); [0073]ggt (gamma-glutamyl peptidase; SEQ ID NO. 51, 52); [0074]yqjN (unknown function--similar to the amino acid decomposition; SEQ ID NO. 1, 2); [0075]yppF (unknown function; SEQ ID NO. 77, 78); [0076]gene (homolog to SEQ ID NO. 53, 54) coding for a hypothetical protein of unknown function; [0077]gene (homolog to SEQ ID NO. 41, 42) coding for a hypothetical protein of unknown function; [0078]ycnl (unknown function--similar to proteins of unknown function; SEQ ID NO. 27, 28); [0079]htrA (serine protease Do; Heat shock protein; SEQ ID NO. 7, 8); [0080]gene (homolog to SEQ ID NO. 91, 92) coding for a putative protein (putative ABC transporter/amino acid permease); [0081]citA (auxiliary citrate synthase I; SEQ ID NO. 59, 60); [0082]kdgR (transcription repressor of the pectin decomposition operon; SEQ ID NO. 25, 26).

[0083]From further experiments (see Example 5) it was determined that the following gene is also inventively suitable as an indicator: [0084]tnrA (a global, transcription regulator involved in the nitrogen metabolic regulation; SEQ ID NO. 99, 100).

[0085]A solution to the posed problem therefore consists of a nucleic acid-binding chip provided with probes for at least three of the following 50 genes: kdgR, citA, gene coding for a putative protein (putative ABC transporter/amino acid permease) (homolog to SEQ ID NO. 91), htrA, ycnl, gene coding for a hypothetical protein of unknown function (homolog to SEQ ID NO. 41), gene coding for a hypothetical protein of unknown function (homolog to SEQ ID NO. 53), yppF, yqjN, ggt, alsR, glnA, nrgA, yciC, yvtA, nrgB, gene coding for a conserved hypothetical protein of unknown function (homolog to SEQ ID NO. 19), gene coding for a putative protein (ATP-binding protein of a putative ABC-transporter) (homolog to SEQ ID NO. 55), ycnJ, glnR, yvlA, yncE, yvlB, gene coding for a putative protein (putative hydrolase) (homolog to SEQ ID NO. 47), trpF, ydfS, trpD, gene coding for a putative protein (putative phage capsid protein) (homolog to SEQ ID NO. 21), ycnK, trpB, trpC, gene coding for a putative protein (putative transcription regulator) (homolog to SEQ ID NO. 17), gene coding for a putative protein (putative serine protease) (homolog to SEQ ID NO. 9), gene coding for a putative protein (putative glycosyl hydrolase/lysozyme) (homolog to SEQ ID NO. 85), gene coding for a putative protein (putative malate synthase, EC 4.1.3.2) (homolog to SEQ ID NO. 45), gene coding for a putative protein (putative Na(+)-bonded D-alanine-glycine permease) (homolog to SEQ ID NO. 49), gene coding for a hypothetical protein of unknown function (homolog to SEQ ID NO. 95), nasD, gene coding for a putative protein (putative ammonium transporter) (homolog to SEQ ID NO. 63), ycdH, nasC, gene coding for a hypothetical protein (close homolog to the aldehyde dehydrogenase DhaS) (homolog to SEQ ID NO. 15), nasB, trpE, pckA, gene coding for a putative protein (putative nitrogen regulation protein P-II) (homolog to SEQ ID NO. 93), nasF, yrkC, gene coding for a hypothetical protein of unknown function (homolog to SEQ ID NO. 81), tnrA,

wherein the total number of all the different probes specific for the nitrogen metabolism is not greater than 80.

[0086]More detailed information on these genes can be found in Examples 1 to 3 and the Tables 1 to 3 of the present application. These genes as obtainable from B. licheniformis DSM 13 are disclosed in the sequence listing. Most of them are also described from other species (see below). Some of them, however, concern putative (assumed/probable) genes or genes that code for putative (assumed/probable) enzymes. According to the invention, these are defined as far as possible, based on databank comparisons, as those with a putative (assumed) function and in addition as homologs to the found B. licheniformis genes. On this, two different points must be explained:

[0087]Firstly, for one or others, it could emerge from a biochemical analysis that these putative functions are not consistent with the real function. In this case the putative function will not be retained. In fact, these results do not challenge the invention in so far as according to the invention, it solely comes down to the observation of increased transcription in connection with the nitrogen deficiency, such that the gene activity in question, independently of the initially performed enzyme activity, can for all intents and purposes serve as the indicator for nitrogen deficiency.

[0088]secondly, in the absence of a gene name, no matching definition can be found for this other than that about the gene itself. One therefore speaks of homologs. Thus one assumes that in species other than B. Licheniformis under nitrogen deficiency the homologous genes are activated. Should it turn out that in a species under consideration a plurality of homologs exists to one of these genes, and said homologs are capable of transcript formation in vivo, then the indication "homolog to SEQ ID NO. . . . ) refers to the closest similarity of each of the various genes under consideration.

[0089]According to the invention, at least three of these genes are selected so as to obtain the conclusion with the highest possible reliability, i.e. in order to eliminate a single false positive signal traced back to only one probe type.

[0090]According to the invention, a nucleic acid-binding chip is understood to mean all objects that are provided with nucleic acid-specific probes and when binding to one or more specifically recognized nucleic acids deliver an analysable signal.

[0091]The composition of chips that are doped with nucleic acids as probes is known from the prior art presented in the introduction. In principle they can all be used for embodiments of the present invention. They are based on the principle of the nucleic acid hybridisation of the mRNA to be detected (or a molecule derived therefrom) with the probe supplied on the chip. According to the system for evaluating the signal initiated by the hybridisation, one distinguishes between chips having an optical and an electrical analysis system. According to the invention, both systems can be used in principle.

[0092]Such chips are employed as follows for monitoring each of the bioprocesses under consideration: At a defined time a sample with the biological material to be analysed is taken from the process. RNA, especially mRNA, is isolated from the sample by known methods, for example by cell digestion and the use of a denaturing buffer. The RNA is self tagged or used as the starting molecule for a molecule introduced into the measurement (for example cDNA obtained by reverse transcription) and the resulting molecules in a buffer are preferably conducted over/through the chip. During hybridisation (sandwich marking) of a prepared RNA or its derivative with the homologous (i.e. congruent with respect to its sequence) probes provided on the chip (target nucleic acid, for example target DNA or target nucleic acid analog) there results a corresponding optically or electronically analysable signal. This is based for example on the marking of the binding mRNA or a transcript thereof with a color marker or fluorescent marker, a hybridisation with a second probe or on a secondary detection reaction, for example via a RT-PCR.

[0093]As at any one time a plurality of molecules is generally bound to the chip from the same probe, the strength of the hybridisation signal over a given--in the individual case to be optimised if needed--area is proportional to the number of specific mRNA present in the sample at the time of sampling. In this way the strength of the signal is a direct measure of the activity of the gene in question at the time of sampling.

[0094]The time span between sampling and measurement should be as short as possible, for example by a largely automated sampling, its preparation and passage over/through the sensor.

[0095]In principle, all plants, animals and microorganisms, especially those that are commercially utilized, can be considered as the organisms monitored using an inventive chip. From the application DE 19860313 A1 entitled "Process for the recognition and characterization of agents against plant pathogens", it emerges that metabolic situations must be observed in plants, particularly useful plants. Similarly, livestock or laboratory animals, for example can also be observed. Eukaryotic cell cultures are of quite considerable commercial interest, for example for the production of monoclonal antibodies, and in particular fermentative production of food, for example via alcoholic fermentation carried out by yeasts. Bacteria are utilized, in particular, for industrial production of proteins or low molecular weight desired substances (biotransformation), for example vitamins or antibiotics.

[0096]According to the invention, probes are understood to mean all molecules that are capable of incurring an essentially specific interaction with nucleic acids (to bind them). According to the invention, this interaction is exploited in order to obtain, in the context of a suitable configuration (chip), a largely unambiguously assignable and analysable signal.

[0097]From the chemical point of view, an inventive probe is a mostly a compound which is capable of binding mRNA molecules or nucleic acids derived therefrom through hydrogen bonds, as is the case, for example, for the interaction of the two strands of a DNA or for DNA-RNA interaction. For example, this can be a DNA, which is more stable to hydrolysis than RNA.

[0098]In addition, further molecules are known in the prior art, in particular chemically synthesized ones, which biomimetically make possible the same interaction but are more stable than DNA, for example by exchanging the phosphate ester bonds of the backbone against less hydrolysis-sensitive bonds. Such nucleic acid-analog probes characterize preferred embodiments of the present application (see below). The corresponding specific probes would have to be synthesized according to the example of the sequence listing related to this application. Advantageously, it should be possible to use chips of the invention multiple times, in particular during a single observed process in the course of which continuous monitoring is desirable.

[0099]Limiting the usability of a probe is thus in each case the extent of homology between the probe provided and the mRNA or the nucleic acid derived therefrom respectively, which is to be recognized by way of hybridization. Ultimately, the extent of hybridization of the probe with the mRNA to be detected (see above) decides its usability as a probe and, in individual cases, has to be optimized experimentally and/or taken into account by way of adjusting signal evaluation. Under the conditions determined by the construction of the measuring apparatus and other influences, a hybridization must take place which can specifically be attributed only to the gene of interest, is sufficiently strong in order to give a positive signal and, on the other hand, is not too strong such that, after the signal has been generated, the mRNA diffuses off in order to render the binding site empty for the next molecule, or to enable the signal to decay; the last, if needed in the course of a suitable washing step.

[0100]It is, however, necessary to estimate the extent of homology between the genes in question prior to using inventive chips for an organism of interest; if the affinity of the mRNAs to be detected to the initially introduced probes is not sufficient, to anchor those probes by way of the same genes of the invention from closer related species on the chip and to carry out calibration measurements in order to obtain reliable information as to which signal strength corresponds to which concentration of special mRNA.

[0101]The identification of the important 50 genes for the present invention is described in the examples of the present application. Their sequences obtained from B. licheniformis are presented in the sequence listing of the present application (SEQ ID NO. 1 to 100), wherein the sequences with odd numbers concern DNA sequences and each sequence with one higher number concern the amino acid sequences derived therefrom. Whereas the DNA sequences can be directly used for manufacturing probes (see above), the amino acid sequences serve for example to verify the gene function using sequence databank comparisons and moreover can be used to generate similar nucleic acid recognizing probes using back translations of the genetic code.

[0102]As is illustrated in example 1, numerous different gene transcripts, i.e. mRNA molecules were investigated, especially those that were known to participate in the nitrogen metabolism. These mRNA molecules were isolated at various times during the transition of B. licheniformis DSM 13 to a state of nitrogen deficiency. Example 1 likewise describes the way in which the increase in concentration of said mRNA inside B. licheniformis cells was determined experimentally. Possible alternative determinations of this may be established in the prior art; the compilation in Table 1 (Example 2) is critical to the understanding of the present invention. It depicts the changes in the concentrations of a total of 210 mRNAs linked to the transition. In this connection, the following thresholds of the ratio of the amount of RNA of the particular gene to the control value were considered significant: according to the invention, the genes whose RNA has a ratio of >3 (i.e. at least a three-fold increase) are considered induced; a distinct induction is present at a ratio of >10; genes having an RNA ratio of <0.3 (i.e. a decrease to less than 30%) are distinctly repressed. For the 210 genes listed in Table 1, at least a three-fold increase was observed at any of the observed times.

[0103]As is proven in Example 3, among these 210 genes there are only 67 genes having an at least 8 fold induction at any of the observed times under the conditions of nitrogen deficiency described in Example 1. Among these, 18 can be regarded as being not particularly specific, such that surprisingly solely the 49 genes listed in Table 2 remain as specifically distinctly induced genes under nitrogen deficiency. According to the invention, these 49 genes are considered representative indicators of a nitrogen deficiency condition. Further information about these genes, for example about their function or deviating start codons, can be found in Tables 2 and 3 and in the sequence listing; the particular English names for the corresponding proteins have already been listed above, in the sequence of data presented in Table 3.

[0104]According to Example 5, the separately identified gene tnrA shows a more than 15 fold induction 30 minutes after the transition into the stationary phase induced by nitrogen deficiency, such that this gene can also be utilized for the realization of the inventive teaching and is correspondingly preferred.

[0105]All these genes have been described in each case individually in the prior art. They can be found for the various organisms in generally accessible databases. As mentioned above, the sequences indicated in the sequence listing for B. licheniformis DSM 13 have been determined from this microorganism and are virtually identical to the information described in the publication "The Complete Genome Sequence of Bacillus licheniformis DSM13, an Organism with Great Industrial Potential" (2004) by B. Veith et al. in J. Mol. Microbiol. Biotechnol., Volume 7(4), pages 204 to 211 and additionally accessible under the entry AE017333 (bases 1 to 4 222 645) in the GenBank database (see above). The B. licheniformis DSM 13 strain is generally obtainable in the Deutsche Sammlung von Mikroorganismen and Zellkulturen GmbH, Mascheroder Weg 1b, 38124 Braunschweig, Germany (htt://www.dsmz.de). It has the deposition number ATCC 14580 at the American Type Culture Collection, 10801 University Boulevard, Manassas, Va. 20110-2209, USA (http;//www.atcc.org).

[0106]A large proportion of the genes from other organisms, which correspond to the 50 genes mentioned, have likewise been deposited in generally accessible databases, for example for the well-characterized species B. subtilis and E. coli which are generally regarded as model organisms of Gram-positive and Gram-negative bacteria, respectively. The corresponding sequences may be found, for example, in the databases of Institut Pasteur, 25, 28 rue du Docteur Roux, 75724 Paris CEDEX 15, France, which are accessible via the internet addresses http://genolist.pasteur.fr/Colibri/ (for E. coli) and http://genolist.pasteur.fr/Colibri/ (for B. subtilis), respectively (as of 12.2.2004). Other databases suitable for this are that of the EMBL-European Bioinformatics Institute (EBI) in Cambridge, United Kingdom (http://www.ebi.ac.uk), Swiss-Prot (Geneva Bioinformatics (GeneBio) S.A., Geneva, Switzerland (http://www.genebio.com/sprot.html) or GenBank (National Center for Biotechnology Information NCBI, National Institutes of Health, Bethesda, Md., USA).

[0107]Said "corresponding genes" mean those which in each case code for the proteins which catalyze the same chemical reaction in the observed organism or which are involved in the same physiological process as the cited 50 proteins in B. licheniformis DSM 13. For other organisms, most of these have similar names and abbreviations to those indicated for B. licheniformis in Tables 1, 2 and 3 because these names represent the particular function. If in doubt, they can be recognized via their sequence, which in each case is the next similar (most homologous) for the organism in question to the sequences mentioned herein. When assigning the function, the similarity of the amino acid sequences to one another is especially decisive because the amino acids are the carriers of the function of the protein and, owing to the degeneracy of the genetic code, various nucleotide sequences can code for the same amino acid sequence.

[0108]Particularly high degrees of relationship exist between closely related species. Thus it can be assumed in principle that for most of the cited 50 genes, homologs can be found in all species, even in cyanobacteria, in eukaryotic cells such as, for example, fungi, or in Gram-negative species such as E. coli or Klebsiella. This probability is even higher for Gram-positive bacteria, in particular of the genus Bacillus, because B. licheniformis DSM 13, from which the sequences listed in the sequence listing are derived, is such a Gram-positive bacterium. It can moreover be assumed that in increasingly related organisms the homologous genes are also increasingly subjected to the same regulatory mechanisms or regulatory mechanisms acting in the same way; therefore, these homologs should also indicate the same metabolic situation, in particular a nitrogen deficiency. In this respect, B. licheniformis is a good choice of an exemplary organism because the commercially likewise particularly important species B. subtilis, B. amyloliquefaciens, B. lentus, B. globigii are likewise Bacilli and therefore Gram-positive. This is in accordance with the concerned aspect of the stated object.

[0109]It should be noted that, in order to practice the invention for a particular species, not all of the 50 mentioned genes must be known but that only a few of them (see below) are sufficient in order to reproduce nitrogen metabolism and, in particular, to be able to detect the transition to a nitrogen deficiency condition. Nevertheless, the reliability of the information about the nitrogen supply state increases with an increasing number of probes. If a plurality of genes are known that on the basis of the present disclosure appear to be suitable in principle, it is recommended to carry out an expression study prior to preparing a corresponding chip, in order to check whether the genes in question actually allow significant information, similarly to the representation in Example 1 or else via a Northern analysis. Shifts regarding the level of expression caused by nitrogen deficiency may be more likely the less related the observed species is to B. licheniformis, such that subgroups (where appropriate other subgroups than the ones listed below) of these 50 genes prove to be particularly suitable and therefore preferred.

[0110]Accordingly, the preparation of an inventive nucleic acid-binding chip for an organism not specified herein must involve the identification of the homologous genes that correspond to at least some of the genes specified for B. licheniformis, for example by comparing the known DNA sequences of the organism in question with the sequences indicated herein. These or parts thereof (see below) may then serve per se as probes or as a template for synthesizing corresponding probes that are applied to a nucleic acid-binding chip by methods known per se.

[0111]If it is the case that some homologous sequences have not been deposited in databases, it is possible for the person skilled in the art to synthesize particular probes on the basis of the sequences disclosed in the sequence listing of the present application in order to screen with the aid thereof a gene library generated for the desired organism (genomic or preferably on the basis of the cDNA) for the homolog in question by generally standard methods. As an alternative to this, it is also possible, on the basis of the DNA sequences indicated in the sequence listing, to synthesize oligonucleotides that serve as PCR primers so as to amplify the genes in question or parts thereof that can be used as probes from a DNA preparation of the complete genome or from a cDNA preparation of the organism of interest. These or parts thereof (see below) can be employed as probes on inventive nucleic acid-specific chips.

[0112]An essential feature of the present invention is the fact that the total number of all the different nitrogen metabolism-specific probes does not exceed 80. This feature correlates with the stated object, according to which it should mainly focus on those nucleic acid-binding chips whose number of occupiable places is comparatively low due to their type of construction. In particular, these are the electrically analyzable chips.

[0113]Further nitrogen metabolism-specific probes may include, for example, those which are induced by an excess of nitrogen, and may include also others which appear not to be directly associated with nitrogen metabolism, but which may be defined as such due to said inducibility. As a result, such a chip also produces analyzable information useful in the observed process, after nitrogen deficiency has been overcome, for example by carrying out appropriate countermeasures.

[0114]Furthermore, nucleic acid-specific probes are usually in each case only fragments of the complete genes (see below). In individual cases, for example with regulation via splicing or with large, multifunctional polypeptides, it may therefore be sensible to detect the same gene with two or more different probes. Corresponding embodiments are therefore, where appropriate, characterized by more than 50 probes which, however, do not respond to more than said 50 genes.

[0115]Depending on the process to be observed, probes for additional genes or gene products may also be comprised on the inventive chips (see below).

[0116]On the other hand, the core of the invention consists specifically of the specificity of the chip in question, with which a special metabolic situation is to be recorded. In the case of such a specific question, the preparation of a chip with more than 80 probes responding to various genes, or even of a chip that reproduces a large part of the genome of an organism, is not part of the invention described herein, due to the complexity associated therewith. Rather, both kinds of chips can be sensibly used in parallel in an observed bioprocess: thus the chips containing a number of various gene probes or containing a representative cross section of various, possibly concerned situations, as are provided by the application WO 2004/027092 A2, can provide a rough overview of the condition of the organism in question, whereas a chip of the invention is used as a control, should there be reason to be concerned that the cells in question could enter a nitrogen deficiency state.

[0117]A preferred embodiment is an inventive nucleic acid-binding chip that is doped increasingly preferably with the probes indicated above in the order indicated there.

[0118]This means the genes listed in Table 3 in reverse order. Accordingly, chips with probes for the kdgR gene (transcription repressor of the pectin break down operon; SEQ ID No. 25, 26) are least preferred among the inventive chips with respect to gene selection for the formation of probes, since this gene has the weakest induction among the mentioned 50 genes, whose transcription is enhanced in a significant manner due to nitrogen deficiency. In contrast, those chips with a probe for the gene that codes for a hypothetical protein of unknown function; (homologous to SEQ ID No. 81, 82) are most preferred in regard to gene selection. This is because this gene showed a more than 146 fold induction over the starting level and therefore has the highest induction of all the genes measured. The signal linked thereto should thus be the most suitable of all the investigated genes for indicating the metabolic situation "nitrogen deficiency".

[0119]According to Example 5, tnrA is between trpC and trpB and is correspondingly preferred.

[0120]A preferred embodiment is an inventive nucleic acid-binding chip, wherein at least one, increasingly preferably two or three of the probes are selected from the following 34 genes:

gene coding for a conserved hypothetical protein of unknown function (homolog to SEQ ID NO. 19), gene coding for a putative protein (ATP-binding protein of a putative ABC-transporter) (homolog to SEQ ID NO. 55), ycnJ, glnR, yvlA, yncE, yvlB, gene coding for a putative protein (putative hydrolase) (homolog to SEQ ID NO. 47), trpF, ydfS, trpD, gene coding for a putative protein (putative phage capsid protein) (homolog to SEQ ID NO. 21), ycnK, trpB, trpC, gene coding for a putative protein (putative transcription regulator) (homolog to SEQ ID NO. 17), gene coding for a putative protein (putative serine protease) (homolog to SEQ ID NO. 9), gene coding for a putative protein (putative glycosyl hydrolase/lysozyme) (homolog to SEQ ID NO. 85), gene coding for a putative protein (putative malate synthase, EC 4.1.3.2) (homolog to SEQ ID NO. 45), gene coding for a putative protein (putative Na(+)-bonded D-alanine-glycine permease) (homolog to SEQ ID NO. 49), gene coding for a hypothetical protein of unknown function (homolog to SEQ ID NO. 95), nasD, gene coding for a putative protein (putative ammonium transporter) (homolog to SEQ ID NO. 63), ycdH, nasC, gene coding for a hypothetical protein (close homolog to the aldehyde dehydrogenase DhaS) (homolog to SEQ ID NO. 15), nasB, trpE, pckA, gene coding for a putative protein (putative nitrogen regulation protein P-II) (homolog to SEQ ID NO. 93), nasF, yrkC, gene coding for a hypothetical protein of unknown function (homolog to SEQ ID NO. 81), tnrA,

[0121]In the studies carried out in the examples on the basis of B. licheniformis DSM 13 and depicted in Examples 1 to 3 and 5, these genes exhibited gene inductions that were elevated by at least a factor of 10. Correspondingly preferred embodiments are thus characterized by the first 33 of the genes listed in Table 3.

[0122]Further preferred are those inventive nucleic acid-binding chips, wherein at least one, increasingly preferably two or three of the probes are selected from the following 20 genes:

trpC, gene coding for a putative protein (putative transcription regulator) (homolog to SEQ ID NO. 17), gene coding for a putative protein (putative serine protease) (homolog to SEQ ID NO. 9), gene coding for a putative protein (putative glycosyl hydrolase/lysozyme) (homolog to SEQ ID NO. 85), gene coding for a putative protein (putative malate synthase, EC 4.1.3.2) (homolog to SEQ ID NO. 45), gene coding for a putative protein (putative Na(+)-bonded D-alanine-glycine permease) (homolog to SEQ ID NO. 49), gene coding for a hypothetical protein of unknown function (homolog to SEQ ID NO. 95), nasD, gene coding for a putative protein (putative ammonium transporter) (homolog to SEQ ID NO. 63), ycdH, nasC, gene coding for a hypothetical protein (close homolog to the aldehyde dehydrogenase DhaS) (homolog to SEQ ID NO. 15), nasB, trpE, pckA, gene coding for a putative protein (putative nitrogen regulation protein P-II) (homolog to SEQ ID NO. 93), nasF, yrkC, gene coding for a hypothetical protein of unknown function (homolog to SEQ ID NO. 81), tnrA,

[0123]In the studies carried out in the examples on the basis of B. licheniformis DSM 13 and depicted in Examples 1 to 3 and 5, these genes exhibited gene inductions that were elevated by at least a factor of 15.

[0124]Further preferred are those inventive nucleic acid-binding chips, wherein at least one, increasingly preferably two or three of the probes are selected from the following 12 genes:

nasD, gene coding for a putative protein (putative ammonium transporter) (homolog to SEQ ID NO. 63), ycdH, nasC, gene coding for a hypothetical protein (close homolog to the aldehyde dehydrogenase DhaS) (homolog to SEQ ID NO. 15), nasB, trpE, pckA, gene coding for a putative protein (putative nitrogen regulation protein P-II) (homolog to SEQ ID NO. 93), nasF, yrkC, gene coding for a hypothetical protein of unknown function (homolog to SEQ ID NO. 81),

[0125]In the studies carried out in the examples on the basis of B. licheniformis DSM 13 and depicted in Examples 1 to 3, these genes exhibited gene inductions that were elevated by at least a factor of 20.

[0126]Further preferred are those inventive nucleic acid-binding chips, wherein at least one, increasingly preferably two or three of the probes are selected from the following 4 genes:

gene coding for a putative protein (putative nitrogen regulation protein P-II) (homolog to SEQ ID NO. 93), nasF, yrkC, gene coding for a hypothetical protein of unknown function (homolog to SEQ ID NO. 81).

[0127]In the studies carried out in the examples on the basis of B. licheniformis DSM 13 and depicted in Examples 1 to 3, these genes exhibited gene inductions which were elevated by at least a factor of 30, in the case of yrkC by more than 60 and in the last case even significantly more than a factor of 100.

[0128]In preferred embodiments, nucleic acid-binding chips of the invention are doped with at least 4, 5, 6, 7, 8, 9, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 35, 40, 45 or 50 of the probes specified for the present invention.

[0129]The more of these probes that respond to a corresponding signal, the more reliable is the information related thereto about the supply or inadequate supply of nitrogen at that moment. Thus it is also sensible to combine the particularly meaningful probes which therefore characterize preferred embodiments with apparently less meaningful ones in order to be able to rule out false-positive signals. A further advantage consists of combining, according to the information in Table 2, those probes with one another that produce signals of different strength at different times indicated there. Thus it is possible to estimate the time interval from the onset of nitrogen deficiency to the sampling and the possibility of said deficiency being attributed to a particular environmental influence--while recording the culturing conditions.

[0130]With respect to their specific sequences, various probes which, however, respond to the same mRNA, for example fragments which hybridize with different regions of the same mRNA (see below), may be present several times in order to increase the read-out reliability, but they are only counted once for the purposes of this aspect of the invention, since they respond, in the best case equally strongly to the same signal and would in principle be exchangeable with one another.

[0131]In preferred embodiments of nucleic acid-binding chips of the invention, the total number of all the different probes does not exceed, in increasing preference 80, 75, 70, 65, 60, 55, 50, 40, 30, 20 or 10.

[0132]This corresponds to the inventive concept stated above, according to which the chips described herein are intended to monitor a special metabolic aspect so that it is not necessary to apply a larger number of probes to the chips in question than is required for recording this situation. This does not affect the situation in which it may be sensible in individual cases to employ more than one probe for detecting the same mRNA and/or to apply individual probes that are linked to the production of a valuable substance of interest. Overall, the present invention moves within the specified scope in order to be able to include within the scope of protection chips that for technical reasons have only a few binding sites.

[0133]In preferred embodiments of nucleic acid-binding chips of the invention, the probes referred to as being relevant to the invention are those which react to the concerned, respectively most homologous, in vivo transcribable genes from the organism chosen for the bioprocess, preferably those which are derived from the concerned, respectively most homologous, in vivo-transcribable genes of this same organism.

[0134]To this end, it has already been stated above that the sequences disclosed in the present application have been obtained from B. licheniformis and should be suitable particularly for monitoring related species, in particular those of the genus Bacillus, owing to the generally known relationships. This applies both to the identified genes to which it was possible to assign specific functions based on database comparisons and which should in principle encode the same function in other species, and to those which have been defined only by their sequences. To this end, the most closely related genes in the observed organism are used according to the invention for deriving corresponding probes. However, care must be taken here that probes are generated not for pseudo genes but for those that are actually transcribed to mRNA under in vivo conditions, i.e. which result in a nucleic acid signal that can be measured in the cytoplasm.

[0135]From a statistical point of view, however, such a chip should be all the more successfully usable, the better is the interaction of the chosen probes with the nucleic acids being measured. As a result, especially with a decreasing degree of relationship to B. licheniformis, it becomes increasingly necessary not to rely on the indicated sequences for this hybridization, but--if sequence differences exist--to use those for the homologous genes from the species in question. The latter sequences can, as explained above, be obtained by methods per se, in particular gene library screening or PCR with primers based on the sequences disclosed herein (where appropriate in the form of "mismatch primers" containing certain, random sequence variations).

[0136]In preferred embodiments of nucleic acid-binding chips of the invention, the organism selected for the bioprocess is a representative of unicellular eukaryotes, Gram-positive or Gram-negative bacteria.

[0137]This is because these groups include the commercially most employed organisms, in particular if the bioprocess to be observed is a fermentation. This includes, for example, fermentative processes, for example for producing wine or beer, or biotechnological production of valuable substances such as proteins or low molecular-weight compounds.

[0138]Various organisms are chosen for a biotechnological method, depending on the type of the desired product. In the context of the invention, this means not only the producer strains but also any organisms upstream of the production process, for example for the cloning of corresponding genes or the selection of suitable expression vectors. In this connection, the need for recording nitrogen limitation is in principle present during each part of the process.

[0139]In preferred embodiments of nucleic acid-binding chips of the invention, the unicellular eukaryotes are protozoa or fungi, among these in particular yeast, quite particularly Saccharomyces or Schizosaccharomyces.

[0140]This is because the latter are employed intensively as host cells, in particular for the gene products of eukaryotes, in addition to being employed for producing alcoholic beverages and other food obtained by fermentation. The former usage is particularly advantageous, if said gene products are to undergo special modifications that can only be carried out by these strains, such as glycosylations of proteins, for example.

[0141]This subject matter also includes inventive chips that are geared to monitor the course, in particular the growth, of cell cultures of higher eukaryotes, for example of rodents or humans. In a certain sense, they may likewise be understood as meaning, at least substantially, unicellular eukaryotes that are of considerable commercial importance, in particular in immunology, for example for producing monoclonal antibodies.

[0142]In preferred embodiments of nucleic acid-binding chips of the invention, the Gram-positive bacteria are Coryneform bacteria or those of the genera Staphylococcus, Corynebacteria or Bacillus, in particular of the species Staphylococcus carnosus, Corynebacterium glutamicum, Bacillus subtilis, B. licheniformis, B. amyloliquefaciens, B. agaradherens, B. stearothermophilus, B. globigii or B. lentus, and quite particularly B. licheniformis.

[0143]This is because they are industrially particularly important producer strains. They are employed in particular for producing low molecular weight chemical compounds, for example vitamins or antibiotics, or for producing proteins, in particular enzymes. Particular mention must be made here of amylases, cellulases, lipases, oxidoreductases and proteases. The particular orientation toward B. licheniforms can be explained by the fact that the sequences indicated in the sequence listing have been obtained from this species and that, as described in Examples 2 and 3, it was possible to prove their connection with the transition to nitrogen deficiency.

[0144]In no less preferred embodiments of nucleic acid-binding chips of the invention, the Gram-negative bacteria are those of the genera E. coli and Klebsiella, in particular derivatives of Escherichia coli K1 2, of Escherichia coli B or Klebsiella planticola, and quite particularly derivatives of the strains Escherichia coli BL21 (DE3), E. coli RV308, E. coli DH5α, E. coli JM109, E. coli XL-1 or Klebsiella planticola (Rf).

[0145]This is because they are used for producing biological valuable substances, both on the laboratory scale, for example cloning and expression analysis, as well as on the industrial scale.

[0146]In preferred embodiments of nucleic acid-binding acids of the invention, at least one, increasingly preferably a plurality, of the probes specified in connection with the invention described herein is/are derived from the sequences listed in the sequence listing under numbers SEQ ID No. 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97 and 99.

[0147]This is because it was possible to prove the connection of these genes with the transition to nitrogen deficiency in B. licheniformis, as described in Examples 2 and 3. These sequences should therefore be used in particular, if B. licheniformis or other, especially related, Bacillus species are to be monitored.

[0148]Preferred embodiments of nucleic acid-binding chips of the invention are those which are additionally doped with at least one probe for an additional gene, in particular one which is metabolically associated with the gene(s) additionally expressed depending on the process, very particularly for one of these or this one itself.

[0149]As explained above, the observed processes are of an industrial interest that is often related to further specific genes. This is, for example if a protein is to be produced, the gene for this protein and, if a low molecular-weight compound is to be produced, one or more gene products which are on the synthetic pathway of the compound in question or which regulate said pathway. Other genes intrinsic to the cell may also be affected, for example metabolic genes which must be increasingly produced in the course of product production, for example an oxidoreductase intrinsic to the cell, if the product is to be obtained from a reactant or an intermediate by oxidation or reduction.

[0150]Moreover, in certain biological processes, in particular the production of commercially relevant compounds by microorganisms, strains geared to the process in question rather than wild type strains are usually employed. They include, apart from transformation with the genes responsible for actual production of the product, provision with selection markers or further metabolic adjustments up to auxotrophies. Such strains have a particular profile of demands on the growth conditions, and some of them possess metabolic genes that are mutated from the wild type genes. Since chips of the invention are advantageously intended to be geared to exactly these strains, quite particularly to the observed bioprocess, these strain-specific peculiarities should be taken into account and may be reflected in the choice of the probes in question.

[0151]In preferred embodiments of such nucleic acid-binding chips of the invention, the gene additionally expressed depending on the process is that for a commercially usable protein, in particular an amylase, cellulose, lipase, oxidoreductase, a hemicellulase or protease, or one which is on a synthetic pathway for a low molecular-weight chemical compound or which at least partially regulates said pathway.

[0152]These are then geared particularly to those bioprocesses, especially fermentations, in which the cited proteins are produced. The latter are commercially particularly important enzymes that are used, for example, in the food industry or detergent industry. The latter case includes in particular the removal of soilings that are hydrolyzable by amylases, cellulases, lipases, hemicellulases and/or proteases, the treatment of the respective materials, in particular by cellulases, or the provision of an enzymatic bleaching system based on an oxidoreductase.

[0153]The latter variant falls within the area of biotransformation, according to which particular, where appropriate additionally introduced, metabolic activities of microorganisms are utilized for the synthesis of chemical compounds.

[0154]In preferred embodiments of nucleic acid-binding chips of the invention, one, preferably a plurality, of the probes specified in connection with the invention described herein is/are provided in single-stranded form, in the form of the codogenic strand.

[0155]This embodiment has the aim of improving hybridization between the probe and the sample to be detected. This applies in particular to the case in which the concerned mRNA content is actually determined from the sample. Since said mRNA is single-stranded and its sequence corresponds to the coding strand of the DNA, optimal hybridization with the complementary, i.e. codogenic, strand should ensue.

[0156]In preferred embodiments of nucleic acid-binding chips of the invention, one, preferably a plurality, of the probes specified in connection with the invention described herein is/are provided in the form of a DNA or of a nucleic acid analog, preferably in the form of a nucleic acid analog.

[0157]This embodiment has the aim of improving the durability and multiple usability of the chips of the invention. This need arises in particular during a single observed process in the course of which constant monitoring is desirable. The durability of chips of the invention, in particular toward nucleic acid-hydrolyzing enzymes, is already increased by providing the probes in the form of a DNA, since the latter is hydrolytically less sensitive per se than an RNA, for example. Even more stable are nucleic acid analogs in which, for example, the phosphate of the sugar-phosphate backbone has been replaced with a chemically different building block that cannot be hydrolyzed, for example, by natural nucleases. Such compounds are known in principle in the prior art and are commercially synthesized by specialized companies for customer indicated sequences. The probes in question can be synthesized, for example, according to the template of the sequences indicated in the sequence listing.

[0158]In preferred embodiments of nucleic acid-binding chips of the invention, one, preferably a plurality of the probes referred to as being relevant to the invention comprises/comprise gene regions that are transcribed into mRNA by the organism to be studied, in particular the gene regions close to the 5' end of said mRNA.

[0159]This takes into account the aspect that in many cases the regulatory DNA sections are also assigned to a special gene. However, the chip of the invention is actually intended to be employed for detecting the mRNA actually present in the observed cells, such that, for the purpose contemplated herein, only the gene section that is actually translated into mRNA is important. Secondly, it must also be considered that introns occur, in particular in eukaryotes, i.e. the coding region is interrupted by sections which are not translated into mRNA. Probes that contain introns should therefore respond to the mRNA in question only poorly or not at all. To implement this aspect, it is advisable to use cDNA sequences rather than genomic DNA sequences, i.e. those sequences that have been obtained on the basis of the actual mRNA.

[0160]Furthermore, detection of an mRNA often does not require hybridization over the entire length of the sequence. The specific probes therefore need to comprise usually only a smaller part of the gene transcribed to mRNA. Advantageous for this is a selection of a region close to the 5' end of the mRNA, since this region is the first to be transcribed to mRNA and therefore the first to be detectable after activation of the gene. This benefits real-time detection.

[0161]In preferred embodiments of nucleic acid-binding chips of the invention, one, preferably a plurality, of the probes referred to as being relevant to the invention reacts/react to fragments of the concerned nucleic acids, in particular to those whose respective mRNA has a low degree of secondary folding, based on the particular total mRNA.

[0162]This is a further aspect in order to optimize hybridization between the probes and the mRNA to be detected. This is because mRNA molecules frequently have a secondary structure, which is based on hybridization of individual mRNA regions with intrinsic, other regions. Thus, for example loop or stem-loop structures are formed. Such regions, however, usually hybridize less readily with other nucleic acid molecules, even if the latter are homologous. Regions of this kind can be calculated quite accurately by computer programs directed thereto (see below). Thus, to implement this aspect, the gene whose activity is desired to be determined for an organism of interest should be analyzed by such a program and, in order to obtain a suitable probe--usually comprising only a subsection (see below)--sections should be used for which only a low degree of mRNA secondary structures is predicted.

[0163]In preferred embodiments of nucleic acid-binding chips of the invention, one, preferably a plurality, of the probes referred to as being relevant to the invention has/have a length of increasingly preferably less than 200, 150, 125 or 100 nucleotides, preferably from 20 to 60 nucleotides, particularly preferably from 45 to 55 nucleotides.

[0164]This is because the probes used for the detection reaction need to comprise only part of the mRNA to be detected, as long as the signal obtainable via them is still specific enough. This specificity, the distinguishability of different mRNAs, sets the lower limit of the length of the probes in question and must, where appropriate, be determined in preliminary experiments.

[0165]The identification of suitable probe lengths and probe regions is known per se to the person skilled in the art and is normally carried out with the aid of specialized software. Examples of such software are the programs Array Designer from Premier Biosoft International, USA, and Vector NTI® Suite, V. 7, obtainable from InforMax, Inc., Bethesda, USA. These software programs also take into account, for example, predefined probe lengths and melting temperatures, in addition to the secondary structures already mentioned.

[0166]In preferred embodiments of nucleic acid-binding chips of the invention, binding of the mRNA to the concerned probe referred to as being relevant to the invention triggers an electric signal.

[0167]The article by J. Wang (Acc. Chem. Res.; ISSN 0001-4842; Rec. Sep. 12, 2001, pp. A-F), mentioned above, discusses the advantages of an electrically analyzable system over an optical system. It also refers to various embodiments of such sensors, which have been developed in the prior art.

[0168]Thus the time interval from sampling to measuring the signal is currently approximately 24 h for optically analyzable chips. With the aid of an electrical system, the required time is currently less than 2 h (cf. FIG. 1). In contrast to this, the number of simultaneously analyzable samples is currently in the two-digit range with electrically analyzable chips, but rapid development gives reason to believe that this order of magnitude may be exceeded soon. The limiting factors in this are the electronic evaluation units for the various signals.

[0169]An example of a method for quantifying mRNA, which is established in the prior art, is RT-PCT. This method is described in the article "Quantification of Bacterial mRNA by One-Step RT-PCR Using the LightCycler System" (2003) by S. Tobisch, T. Koburger, B. Jurgen, S. Leja, M. Hecker and T. Schweder in BIOCHEMICA, Volume 3, pages 5 to 8. In comparison with this, detection via electrochips has another advantage, namely higher reliability of the data, since these have distinctly smaller ranges of fluctuation compared to RT-PCR.

[0170]The preparation of corresponding electronically analyzable chips is described, for example, in the patent applications WO 00/62048 A2, WO 00/67026 A1 and WO 02/41992, whose entire disclosed content is incorporated into the present application.

[0171]The manner in which function electrically readable chips of a particularly preferred embodiment can be described as follows: the gene-specific probes are bound covalently in a manner known per se to magnetic beads located in specifically designed chambers of said chips. The specific hybridization of the corresponding mRNA on the particular beads occurs in this hybridization chamber whose temperature can be controlled and which can be flushed by the solutions in question. The beads are retained in this chamber by a magnet. After hybridization of the RNA samples on the beads-bound to the DNA probes, unbound RNA is removed in a washing step, as a result of which only specific hybrids are still present in the incubation chamber, in fact bound to the magnetic beads.

[0172]After washing, a detection probe labeled by a biotin-extravidin-bound alkaline phosphatase is introduced into the incubation chamber. This probe binds to a second free region of the hybridized mRNA. This hybrid is then washed again and incubated with the alkaline phosphatase substrate, para-aminophenol phosphate (pAPP). The enzymatic reaction in the incubation chamber releases the redox-active product, para-aminophenol (pAP), which is then passed over the Red/Ox electrode on the electrical chip and the signal is transmitted to a potentiostat.

[0173]A system-specific software (for example MCDDE32) reads the obtained data, and the results may be evaluated and displayed on a computer with the aid of a further program (for example Origin).

[0174]This process can of course be varied with regard to both the technical design of the chips and the evaluation. Thus, for example, the detection reaction may be carried out by way of a different reaction, but preferably a redox reaction due to the electrical principle of measurement.

[0175]One achievement of the present invention is to have identified nitrogen metabolism-specific and thus process-critical genes and to have made them accessible to analysis via correspondingly designed biochips. The advantage of chips over conventional detection methods comprises, in addition to the time saved and higher accuracy, the possibility of detecting the activities of a plurality of different genes in the same sample at the same time by providing a plurality of probes on a single support and of said activities resulting in a more solid and more detailed picture, for example with regard to the time at which a nitrogen deficiency started, with the application to a special problem, described herein.

[0176]A separate subject matter of the invention is therefore the simultaneous use of nucleic acid probes or nucleic acid analog probes for at least three of the following 50 genes:

kdgR, citA, gene coding for a putative protein (putative ABC transporter/amino acid permease) (homolog to SEQ ID NO. 91), htrA, ycnl, gene coding for a hypothetical protein of unknown function (homolog to SEQ ID NO. 41), gene coding for a hypothetical protein of unknown function (homolog to SEQ ID NO. 53), yppF, yqjN, ggt, alsR, glnA, nrgA, yciC, yvtA, nrgB, gene coding for a conserved hypothetical protein of unknown function (homolog to SEQ ID NO. 19), gene coding for a putative protein (ATP-binding protein of a putative ABC-transporter) (homolog to SEQ ID NO. 55), ycnJ, glnR, yvlA, yncE, yvlB, gene coding for a putative protein (putative hydrolase) (homolog to SEQ ID NO. 47), trpF, ydfS, trpD, gene coding for a putative protein (putative phage capsid protein) (homolog to SEQ ID NO. 21), ycnK, trpB, trpC, gene coding for a putative protein (putative transcription regulator) (homolog to SEQ ID NO. 17), gene coding for a putative protein (putative serine protease) (homolog to SEQ ID NO. 9), gene coding for a putative protein (putative glycosyl hydrolase/lysozyme) (homolog to SEQ ID NO. 85), gene coding for a putative protein (putative malate synthase, EC 4.1.3.2) (homolog to SEQ ID NO. 45), gene coding for a putative protein (putative Na(+)-bonded D-alanine-glycine permease) (homolog to SEQ ID NO. 49), gene coding for a hypothetical protein of unknown function (homolog to SEQ ID NO. 95), nasD, gene coding for a putative protein (putative ammonium transporter) (homolog to SEQ ID NO. 63), ycdH, nasC, gene coding for a hypothetical protein (close homolog to the aldehyde dehydrogenase DhaS) (homolog to SEQ ID NO. 15), nasB, trpE, pckA, gene coding for a putative protein (putative nitrogen regulation protein P-II) (homolog to SEQ ID NO. 93), nasF, yrkC, gene coding for a hypothetical protein of unknown function (homolog to SEQ ID NO. 81), tnrA, bound to a nucleic acid-binding chip, preferably to the same chip, for determining the physiological state of an organism undergoing a biological process.

[0177]As explained above, these 50 genes are selected so as to deliver a picture of the situation of the nitrogen metabolism of the observed organism, since they are, as described in Examples 1 to 3 and 5, significantly and comparatively specifically induced during transition of the Gram-positive bacterium B. licheniformis to a nitrogen deficiency condition. A comparable statement can also be expected for other organisms, which have the homologous genes or proteins with essentially the same metabolically relevant properties.

[0178]As likewise described in detail above, such gene activities may in principle be determined in various ways, for example by Northern hybridization. However, analysis with the aid of a nucleic acid-binding chip, in particular an above-described chip, enables a plurality of gene activities to be determined very efficiently at the same time and, moreover, very much in real-time. As a result, the metabolic alterations of an organism undergoing a biological process may be observed in real time and, where appropriate, intervened in a regulatory manner.

[0179]The above statements on nucleic acid-binding chips apply to the uses accordingly specified herein of the probes in question.

[0180]Accordingly, preference is given here to those uses, wherein the total number of all the different nitrogen metabolism-specific probes does not exceed 80.

[0181]Accordingly, further preference is given here to those uses, wherein the probes indicated above as inventive are employed with increasing preference in the order indicated above as preferable (inverse to the order in Table 3 including example 5).

[0182]In accordance with the above statements, increasing preference is given to the following of the above-specified uses of nucleic acid probes or nucleic acid analog probes:

use, wherein at least one, increasingly preferably two or three of the nucleic acid- or nucleic acid analog-probes is/are selected from the following 34 genes: gene coding for a conserved hypothetical protein of unknown function (homolog to SEQ ID NO. 19), gene coding for a putative protein (ATP-binding protein of a putative ABC-transporter) (homolog to SEQ ID NO. 55), ycnJ, glnR, yvlA, yncE, yvlB, gene coding for a putative protein (putative hydrolase) (homolog to SEQ ID NO. 47), trpF, ydfS, trpD, gene coding for a putative protein (putative phage capsid protein) (homolog to SEQ ID NO. 21), ycnK, trpB, trpC, gene coding for a putative protein (putative transcription regulator) (homolog to SEQ ID NO. 17), gene coding for a putative protein (putative serine protease) (homolog to SEQ ID NO. 9), gene coding for a putative protein (putative glycosyl hydrolase/lysozyme) (homolog to SEQ ID NO. 85), gene coding for a putative protein (putative malate synthase, EC 4.1.3.2) (homolog to SEQ ID NO. 45), gene coding for a putative protein (putative Na(+)-bonded D-alanine-glycine permease) (homolog to SEQ ID NO. 49), gene coding for a hypothetical protein of unknown function (homolog to SEQ ID NO. 95), nasD, gene coding for a putative protein (putative ammonium transporter) (homolog to SEQ ID NO. 63), ycdH, nasC, gene coding for a hypothetical protein (close homolog to the aldehyde dehydrogenase DhaS) (homolog to SEQ ID NO. 15), nasB, trpE, pckA, gene coding for a putative protein (putative nitrogen regulation protein P-II) (homolog to SEQ ID NO. 93), nasF, yrkC, gene coding for a hypothetical protein of unknown function (homolog to SEQ ID NO. 81), tnrA,

[0183]among these, preferably for genes from the following 20 genes:

trpC, gene coding for a putative protein (putative transcription regulator) (homolog to SEQ ID NO. 17), gene coding for a putative protein (putative serine protease) (homolog to SEQ ID NO. 9), gene coding for a putative protein (putative glycosyl hydrolase/lysozyme) (homolog to SEQ ID NO. 85), gene coding for a putative protein (putative malate synthase, EC 4.1.3.2) (homolog to SEQ ID NO. 45), gene coding for a putative protein (putative Na(+)-bonded D-alanine-glycine permease) (homolog to SEQ ID NO. 49), gene coding for a hypothetical protein of unknown function (homolog to SEQ ID NO. 95), nasD, gene coding for a putative protein (putative ammonium transporter) (homolog to SEQ ID NO. 63), ycdH, nasC, gene coding for a hypothetical protein (close homolog to the aldehyde dehydrogenase DhaS) (homolog to SEQ ID NO. 15), nasB, trpE, pckA, gene coding for a putative protein (putative nitrogen regulation protein P-II) (homolog to SEQ ID NO. 93), nasF, yrkC, gene coding for a hypothetical protein of unknown function (homolog to SEQ ID NO. 81), tnrA,

[0184]among these, particularly preferably for genes from the following 12 genes:

nasD, gene coding for a putative protein (putative ammonium transporter) (homolog to SEQ ID NO. 63), ycdH, nasC, gene coding for a hypothetical protein (close homolog to the aldehyde dehydrogenase DhaS) (homolog to SEQ ID NO. 15), nasB, trpE, pckA, gene coding for a putative protein (putative nitrogen regulation protein P-II) (homolog to SEQ ID NO. 93), nasF, yrkC, gene coding for a hypothetical protein of unknown function (homolog to SEQ ID NO. 81),

[0185]among these, quite particularly preferably for genes from the following 4 genes:

gene coding for a putative protein (putative nitrogen regulation protein P-II) (homolog to SEQ ID NO. 93), nasF, yrkC, gene coding for a hypothetical protein of unknown function (homolog to SEQ ID NO. 81).

[0186]In accordance with the above statements, the inventive uses preferably concern the simultaneous use of at least 4, 5, 6, 7, 8, 9, 10, 12, 14, 16, 18, 20, 22, 24, 26 28, 30, 35, 40, 45 or 50 of the specified probes.

[0187]In accordance with the above statements, preferred uses of the invention are those, wherein the total number of all the different probes does, with increasing preference, not exceed 80, 75, 70, 65, 60, 55, 50, 40, 30, 20 or 10.

[0188]In accordance with the above statements, further preference is given to uses of the invention for determining a change in the nitrogen metabolism of the organism undergoing the biological process, preferably for detecting a nitrogen deficiency condition.

[0189]In accordance with the above statements, further preference is given to uses of the invention, wherein at least one, increasingly preferably a plurality, of the specified probes is/are derived from the sequences listed in the sequence listing under the numbers SEQ ID No. 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97 and 99.

[0190]A separate subject matter of the invention is formed by methods for the determination of the physiological state of an organism under a biological process by the use of a nucleic acid-binding chip of the invention.

[0191]These methods in principle comprise removing samples of the observed organism during said process, without interrupting the latter, and isolating the mRNA from said samples. These compounds or, where appropriate, compounds derived therefrom, such as cDNA, for example, are passed across an above-described nucleic acid-binding chip which is treated taking into account the method steps indicated above--such as, for example, adequate incubation time or washing off non-specifically binding nucleic acids--and is finally delivered to the detection apparatus. A method protocol of this kind is depicted in principle in FIG. 1 on the basis of the example of electrically analyzable chips.

[0192]The above comments on nucleic acid-binding chips apply accordingly to the methods specified herein for the determination of the physiological state of an organism undergoing a biological process.

[0193]Preference is given to methods of the invention, wherein a change in the nitrogen metabolism of the organism undergoing the biological process, preferably a nitrogen deficiency condition, is determined.

[0194]With respect to this field of use, the genes described in the examples and listed above have been selected. Their significant induction, at least in B. licheniformis, is accompanied by the onset of a nitrogen deficiency condition, such that this can be detected particularly reliably by the cited method, and in fact not only in B. Licheniformis but, with increasingly improving prospects of success, also with increasingly related species (see above). Moreover, this metabolic situation represents a critical point in the lifecycle of many microorganisms. Thus, as illustrated in the examples (see below), the particular onset of nitrogen deficiency was always also associated with a transition to the stationary growth phase. Methods, which facilitate the early recognition of this point in time, serve to delay this transition and, in particular with industrially utilized fermentations, prolong the phase of production of a valuable substance, particularly when dealing with a nitrogen-containing valuable product such as for example a protein.

[0195]A further possibility for conducting the method for the cultivation of microorganisms consists of supplying, beside the first (and therefore better) utilized N-source, a further source. In the phase of the initially observed nitrogen deficiency, the concerned cells, in so far as they are capable, turn themselves to using the second N source that is less utilizable for them. With this transition there is generally observed a decrease in the growth rate, which can be detected at an early stage by means of the inventive method.

[0196]In accordance with the above, preference is given to those inventive methods, wherein the organism selected for the bioprocess is a representative of unicellular eukaryotes, Gram-positive or Gram-negative bacteria.

[0197]In accordance with the above statements, preference is given among said methods to those methods of the invention, wherein the unicellular eukaryotes are protozoa or fungi, among these in particular yeast, quite particularly Saccharomyces or Schizosaccharomyces.

[0198]In accordance with the above statements, preference is given here also to those inventive methods, wherein the Gram-positive bacteria are coryneform bacteria or those of the genera Staphylococcus, Corynebacteria and Bacillus, in particular of the species Staphylococcus carnosus, Corynebacterium glutamicum, Bacillus subtilis, B. licheniformis, B. amyloliquefaciens, B. agaradherens, B. stearothermophilus, B. globigii or B. lentus, and quite particularly B. licheniformis.

[0199]In accordance with the above statements, no less preference is given here also to those inventive methods, wherein the Gram-negative bacteria are those of the genera E. coli and Klebsiella, in particular derivatives of Escherichia coli K122, of Escherichia coli B or Klebsiella planticola, and quite particularly derivatives of the strains Escherichia coli BL21 (DE3), E. coli RV308, E. coli DH5α, E. coli JM109, E. coli XL-1 or Klebsiella planticola αRf).

[0200]In accordance with the above statements, preference is given to those methods of the invention, wherein those specified probes are used, which are derived from the SEQ ID Nos. 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97 or 99 indicated in the sequence listing.

[0201]Among these, preference is again given to those methods, which employ the probes specified herein for the above-listed Gram-positive bacteria, in particular B. licheniformis, since these sequences have been isolated from this very organism and can therefore be applied most successfully to this species.

[0202]In accordance with the above statements, preference is given to those inventive methods, wherein the physiological state is determined at various times of the same process, preferably using a plurality of structurally identical nucleic acid-binding chips, particularly preferably of the same nucleic acid-binding chip.

[0203]In accordance with the above statements, preference is furthermore given to those methods of the invention, wherein the process is a fermentation, in particular the fermentative production of a commercially usable product, particularly preferably the production of a protein or of a low molecular weight chemical compound.

[0204]In accordance with the above statements, preference is given among these methods to those methods of the invention, wherein the low molecular weight chemical compound is a natural substance, a food supplement or a pharmaceutically relevant compound.

[0205]In accordance with the above statements, preference is alternatively also given to those methods of the invention, wherein the protein is an enzyme, in particular an enzyme from the group of the α-amylases, proteases, cellulases, lipases, oxidoreductases, peroxidases, laccases, oxidases and hemicellulases.

[0206]A separate subject matter of the invention is also the possible uses of inventive nucleic acid-binding chips, as have been described in detail above, for determining the physiological state of an organism undergoing a biological process.

[0207]The above comments on nucleic acid-binding chips apply accordingly to the uses specified herein for determining the physiological state of an organism undergoing a biological process.

[0208]In accordance with the above statements, preference is given to methods of the invention, wherein a change in the nitrogen metabolism of the organism undergoing the biological process, preferably a nitrogen deficiency condition, is determined.

[0209]In accordance with the above statements, further preference is given to those inventive methods, wherein the organism selected for the bioprocess is a representative of unicellular eukaryotes, Gram-positive or Gram-negative bacteria.

[0210]In accordance with the above statements, such inventive methods among these are preferred, wherein the unicellular eukaryotes are protozoa or fungi, among these in particular yeast, quite particularly Saccharomyces or Schizosaccharomyces.

[0211]In accordance with the above statements, preference is alternatively given here to those inventive methods, wherein the Gram-positive bacteria are coryneform bacteria or those of the genera Staphylococcus, Corynebacteria and Bacillus, in particular of the species Staphylococcus carnosus, Corynebacterium glutamicum, Bacillus subtilis, B. licheniformis, B. amyloliquefaciens, B. agaradherens, B. stearothermophilus, B. globigii or B. lentus, and quite particularly B. licheniformis.

[0212]In accordance with the above statements, no less preference is given here also to those inventive methods, wherein the Gram-negative bacteria are those of the genera E. coli and Klebsiella, in particular derivatives of Escherichia coli K122, of Escherichia coli B or Klebsiella planticola, and quite particularly derivatives of the strains Escherichia coli BL21 (DE3), E. coli RV308, E. coli DH5α, E. coli JM109, E. coli XL-1 or Klebsiella planticola (Rf).

[0213]In accordance with the above statements, preference is given to those methods of the invention, wherein those specified probes are used, which are derived from the SEQ ID Nos. 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97 or 99 indicated in the sequence listing.

[0214]For the reason already specified above, among these uses, preference is given in turn to those uses which employ the probes specified herein for the above-listed Gram-positive bacteria, in particular B. licheniformis.

[0215]In accordance with the above statements, preference is given to those inventive methods, wherein the physiological state is determined at various times of the same process, preferably using a plurality of structurally identical nucleic acid-binding chips, particularly preferably of the same nucleic acid-binding chip.

[0216]In accordance with the above statements, preference is furthermore given to those methods of the invention, wherein the process is a fermentation, in particular the fermentative production of a commercially usable product, particularly preferably the production of a protein or of a low molecular weight chemical compound.

[0217]In accordance with the above statements, preference is given among these methods to those methods of the invention, wherein the low molecular weight chemical compound is a natural substance, a food supplement or a pharmaceutically relevant compound.

[0218]In accordance with the above statements, preference is alternatively also given to those methods of the invention, wherein the protein is an enzyme, in particular an enzyme from the group of the α-amylases, proteases, cellulases, lipases, oxidoreductases, peroxidases, laccases, oxidases and hemicellulases.

[0219]The present invention is additionally illustrated by the following examples.

EXAMPLES

[0220]All molecular-biological work was carried out by standard methods as can be found, for example, in the manual by Fritsch, Sambrook and Maniatis "Molecular cloning: a laboratory manual", Cold Spring Harbour Laboratory Press, New York, 1989, or comparable specialist literature. Enzymes and kits were used according to the instructions of the particular manufacturer.

Example 1

Identification of the Gene Probes by Chip Analyses

Culturing of Bacteria and Isolation of Samples

[0221]Cells of the Bacillus licheniformis DSM13 strain (obtainable from Deutsche Sammlung von Mikroorganismen and Zellkulturen GmbH, Mascheroder Weg 1b, 38124 Braunschweig, Germany; http://www.dsmz.de) were cultured in nitrogen-limited, synthetic Belitsky minimal medium (1 mM final concentration) with constant shaking at 270 rpm and 37° C. This medium has the following composition: (1.) Basic medium (pH 7.5): 0.015 M (NH₄)₂SO₄, 0.008 M MgSO₄×7H₂O, 0.027 M KCl, 0.007 M Na₃citrate×2H₂O, 0.050 M Tris-HCl and 0.009 M glutamic acid; (2.) supplements: 0.2 M KH₂PO₄, 0.039 M L-tryptophan-HCl, 1 M CaCl₂×2H₂O, 0.0005 M FeSO₄×7H₂O, 0.025 M MnSO₄×4H₂O and 20% (w/v) glucose; and as (3.) medium supplementing plan: 0.14 ml KH₂PO₄, 0.2 ml CaCl₂, 0.2 ml FeSO₄, 0.04 ml MnSO₄, 1 ml glucose. All quantities refer to 100 ml of basic medium.

[0222]In the course of establishing the growth profile, the control sample was removed at an optical density at 500 nm (OD₅₀₀) of from 0.4 to 0.5, with another sample ("transition") being removed in the transient growth phase at OD₅₀₀ 0.8 to 0.9. The supplementing plan for the Belitzky minimal medium, indicated above, was set up in such a way that nitrogen deficiency started at OD₅₀₀ 0.8 to 1.0, thereby introducing the stationary phase. Further samples were taken after another 30, 60, 90 and 120 min. Immediately after their removal, an equal volume of Killing buffer (20 mM NaN₃; 20 mM Tris-HCl, pH 7.5; 5 mM MgCl₂) cooled to 0° C. was added to the samples. Immediately thereafter, the cell pellet was removed by centrifugation at 4° C. and 15000 rpm for 5 min, the supernatant was discarded, and the pellet was either frozen at -80° C. or immediately worked up further.

Cell Disruption

[0223]The cells were disrupted using the "Hybaid RiboLyser® Cell Disruptor" (Thermo Electron Corporation, Dreieich, Germany). This method is based on mechanically destroying the cell wall and the cell membrane with the aid of glass beads of approx. 0.1 mm in size (Sartorius BBI, Melsungen, Germany). The cells to be disrupted which had been resuspended in lysis buffer II (3 mM EDTA; 200 mM NaCl) beforehand were introduced together with the glass beads into a glass (strain maintenance) tube. Acidic phenol was also added in order to prevent RNA degradation by RNases. This tube was then clamped inside the RiboLyser, wherein the glass beads collided with the cells under vigorous shaking, thereby causing disruption of the cells.

Isolation of Total RNA

[0224]After cell disruption, the RNA-containing aqueous phase is separated by centrifugation from the protein and cell fragments, chromosomal DNA and the glass beads, and the RNA is isolated therefrom with the aid of the KingFisher mL apparatus (Thermo Electron Corporation, Dreieich, Germany) using the MagNA Pure LC RNA Isolation Kit I (Roche Diagnostics, Penzberg, Germany). This purification is based on the RNA binding to magnetic glass particles in the presence of chaotropic salts that also inactivate RNases. The magnetic particles here serve as a means of transporting the RNA between various reaction vessels filled with binding, washing and elution buffers. The KingFisher mL utilized for this is a kind of pipetting robot that transports the particles with the bound RNA back and forth between the vessels with the aid of magnets and is also utilized for mixing the samples. Finally, the RNA is detached from the magnetic particles and is then present in a purified state.

RNA Quality and Quantity Control

[0225]The purification of RNA is followed by quality and quantity controls. The Agilent Bioanalyzer 2100 apparatus (Agilent Technologies, Berlin, Germany) utilized for this enables the analysis of RNA on lab-on-a-chip scale. Together with the Agilent "RNA 6000 Nano Kit", total RNA is fractionated gel electrophoretically and thereby offers the possibility of examining the quality with regard to partial degradation and contaminations. This involves detecting ribosomal RNA (16 S and 23 S rRNA). If the latter appear as clear bands, the RNA can be assumed not to have been degraded during processing and therefore to be intact and to be able to be introduced into the subsequent examinations. In addition, the exact concentration was also determined here.

Transcriptome Analyses

[0226]Transcriptome analyses were carried out using total genomic B. licheniformis DSM13 DNA microarrays which had been prepared in the usual manner (for example according to WO 95/11995 A1) and which can be analyzed by an optical system. These DNA microarrays contained two copies of virtually each B. licheniformis gene, such that it was possible to analyze two samples in parallel on the same chip and to average the values obtained.

[0227]The principle of the measurement carried out consists of transcribing in vitro the particular mRNA molecules from the sample taken via reverse transcription to DNA, wherein one of the added deoxyribonucleotides carries a dye marker. These labeled molecules are then hybridized with the known probes located at known sites of the chip, and the particular strength of the signal at the sites in question, which can be attributed to the fluorescent marker, is optically recorded. When using two different fluorescent markers for a control and for a sample actually to be studied, which are made to hybridize simultaneously and thereby compete for binding to the presented probe, different color values are obtained which provide a measure of the ratio of the concentration of the control to that of the sample.

[0228]dUTP labeled with the fluorescent dye cyanine 3 or cyanine 5 was chosen for this labeling. Thus, in each case 25 pg of total RNA of the control (OD₅₀₀ 0.4) were labeled with the fluorescent dye cyanine 3 (Amersham Biosciences Europe GmbH, Freiburg, Germany) and 25 μg of total RNA of the particular stress sample (transient phase, 30, 60 and 120 min) were labeled with the fluorescent dye cyanine 5 (GE Healthcare, Freiburg, Germany). Competitive hybridization of the two samples was carried out at 42° C. on the conventional, total genomic B. licheniformis DSM13 DNA microarray for at least 16 hours.

[0229]After washing the array in order to remove unspecific bonds, the array was read out optically with the aid of the ScanArray® Express Laser Scanner (PerkinElmer Life and Analytical Sciences, Rodgau-Jugesheim, Germany). All hybridizations were repeated, with the samples being labeled with the in each case other dye (dye swap method). Quantitative evaluation of the arrays was carried out using the ScanArray® Express software (available from PerkinElmer Life and Analytical Sciences, Rodgau-Jugesheim, Germany) according to the manufacturer's instructions and with standard parameters.

[0230]The arrays were normalized and evaluated with the aid of the Lucidea Score Card controls (GE Healthcare, Freiburg, Germany). With the aid of this "Score Card" which is used for controlling hybridization efficiency and quality, known control DNAs and "spikes" in the form of oligos have been applied to the array according to the manufacturer's instructions and admixed with complementary sequences during hybridization. Thus it was possible to control, after scanning, the success of said hybridization and incorporation of the dyes. The controls should be present in the same amounts in both samples and therefore appear yellow after scanning or have a ratio of 1 between both channels. The spikes are specific for the particular sample and are applied in various dilutions, i.e. they appear red or green after scanning for the particular sample.

[0231]For expression of the genes, averages of the two hybridizations and the particular standard deviations were calculated. For a significant induction or significant repression, the following thresholds of the ratio of the amount of RNA of the particular gene to the control value were observed: genes whose RNA has a ratio of >3 (i.e. at least a three-fold increase) are regarded as induced; genes having an RNA ratio of <0.3 (i.e. a decrease to less than 30%) are distinctly repressed. These results are listed in Example 2.

Example 2

Genes Induced Under Nitrogen Deficiency

[0232]Table 1 below lists all 210 Bacillus licheniformis DSM13 genes determined in Example 1 whose induction (of at least a factor of 3) was observed under the conditions of nitrogen deficiency described in Example 1. The first two columns indicate the particular name of the derived protein and, respectively, its abbreviation (if available); the "BLi number" corresponds to the "locus_tag" of the B. licheniformis complete genome accessible under the entry AE017333 (bases 1 to 4 222 645) in the GenBank database (National Center for Biotechnology Information NCBI, National Institutes of Health, Bethesda, Md., USA; http://www.ncbi.nlm.nih.gov; as of 12.2.2004); this is followed by the factors of increasing the concentration of the in each case corresponding mRNAs, observed at the times indicated at the top.

TABLE-US-00001 TABLE 1 The 210 Bacillus licheniformis DSM13 genes determined in Example 1, whose induction (of at least a factor of 3) under nitrogen deficiency was observed (explanations: see text). Protein name/Gene function Gene BLi--Nr Transition 30 min 60 min 90 min 120 min unknown function - yqjN BLi02550 1.56 2.68 4.48 8.50 8.65 similar to amino acid degradation unknown function gene for a BLi02843 1.03 1.74 2.72 3.36 3.32 hypothetical protein unknown function - yozO BLi02247 1.14 2.67 2.38 3.17 3.05 similar to unknown function proteins aldehyde dhaS BLi02249 1.60 4.31 6.38 5.59 5.99 dehydrogenase bacillopeptidase F bpr BLi01748 1.43 6.26 4.37 7.60 12.43 putative gene for a BLi01747 1.17 3.58 2.49 3.41 3.63 bacillopeptidase F putative protein unknown function - yvyD BLi03774 1.60 10.78 9.72 11.51 8.37 similar to sigma-54 modulating factor of gram-negative bacteria unknown function gene for a BLi00431 1.23 4.74 4.29 4.97 3.47 hypothetical protein nitrogen-regulated PII- nrgB BLi03889 3.61 6.92 5.04 9.99 6.67 like protein ammonium transporter nrgA BLi03888 3.04 5.71 3.91 9.56 7.31 unknown function gene for a BLi00491 2.56 3.79 3.15 3.71 2.75 conserved hypothetical protein unknown function yoeB BLi01308 1.31 22.73 25.86 46.66 39.40 unknown function - yjcG BLi01284 1.23 3.28 2.71 3.07 2.42 similar to unknown function proteins unknown function - ykfA BLi01397 1.86 3.48 4.44 3.15 3.61 similar to immunity to bacteriotoxins dipeptide ABC dppE BLi01396 2.60 8.07 8.32 5.59 6.85 transporter/dipeptide- binding protein (sporulation) dipeptide ABC dppC BLi01394 2.08 4.08 4.62 3.16 3.40 transporter/permease (sporulation) dipeptide ABC dppB BLi01393 2.47 4.19 3.35 2.13 2.46 transporter/permease (sporulation) D-alanyl- dppA BLi01392 2.39 3.78 3.51 2.30 2.64 aminopeptidase pyrroline-5-carboxylate proG BLi01391 1.72 4.49 3.23 1.54 1.72 reductase serine protease Do htrA BLi01390 7.91 8.10 2.03 1.93 1.67 (heat-shock protein) unknown function gene for a BLi01389 4.31 4.00 1.87 1.93 2.03 hypothetical protein unknown function - ypjP BLi02321 1.33 3.12 4.80 3.44 3.93 similar to unknown function proteins unknown function - yplQ BLi02317 1.10 2.44 2.85 3.31 3.10 similar to hemolysin III homolog unknown function - yppQ BLi02302 1.58 4.72 4.17 5.38 3.63 similar to peptide methionine sulfoxide reductase unknown function - yusX BLi03477 1.31 2.92 3.22 3.21 2.95 similar to oligoendopeptidase major intracellular ispA BLi01423 1.11 6.98 4.30 17.19 4.56 serine protease negative regulator of rsiX BLi02448 2.06 4.92 2.62 5.37 5.95 sigma-X activity RNA polymerase ECF- sigX BLi02449 2.24 5.73 2.18 5.40 4.34 type sigma factor anti-sigma F factor (EC spoIIAB BLi02496 2.37 7.05 4.42 4.37 2.02 2.7.1.37)/stage II sporulation protein AB anti-sigma F factor spoIIAA BLi02497 1.43 4.60 3.37 3.17 2.24 antagonist/stage II sporulation protein AA unknown function - ysdB BLi03031 1.42 2.88 2.55 4.02 4.69 similar to unknown function proteins subtilisin carlsberg aprE BLi01109 1.74 13.83 5.92 18.92 7.74 precursor (EC 3.4.21.62) unknown function gene for a BLi00303 1.20 4.92 2.55 5.06 4.40 hypothetical protein unknown function ybdN BLi00302 1.71 4.07 2.95 7.30 5.02 putative serine protease gene for a BLi00301 2.31 13.40 7.51 15.91 11.55 putative protein unknown function - yoaD BLi04062 2.39 1.82 2.44 4.37 4.19 similar to phosphoglycerate dehydrogenase putative transcriptional gene for a BLi04093 1.25 2.63 2.96 3.01 2.80 regulator putative protein citrate synthase III mmgD BLi04094 1.32 5.34 4.06 6.95 3.45 function unknown mmgE BLi04095 0.95 2.34 3.19 3.60 3.10 function unknown function - yqiQ BLi04096 1.11 2.03 3.13 2.77 3.21 similar to phosphoenolpyruvate mutase RNA polymerase ECF- sigW BLi00199 1.04 1.61 3.11 3.58 4.83 type sigma factor unknown function - ybbM BLi00200 1.01 1.79 2.91 3.84 4.17 similar to unknown function proteins unknown function gene for a BLi02331 0.98 1.45 1.93 4.09 3.26 hypothetical protein unknown function gene for a BLi01570 1.33 3.57 1.86 3.01 1.74 hypothetical protein phage shock protein A pspA BLi00640 1.59 2.26 3.17 2.88 3.78 homolog unknown function yfmQ BLi00629 1.38 3.99 2.78 5.91 1.88 transcriptional repressor glnR BLi01992 4.38 11.07 7.61 9.77 8.25 of the glutamine synthetase gene glutamine synthetase glnA BLi01993 5.69 9.51 8.60 9.06 8.61 unknown function yoaW BLi02014 1.16 2.80 3.04 3.22 4.52 unknown function - ydfO BLi02024 1.86 3.64 3.36 3.89 3.07 similar to unknown function proteins unknown function - ywfL BLi03988 2.21 2.33 2.84 5.85 4.75 similar to unknown function proteins putative gene for a BLi03989 2.19 2.76 3.89 6.36 7.16 hydroxybenzoate putative hydroxylase protein putative benzoate gene for a BLi03990 4.12 4.74 6.49 13.68 12.33 transport protein putative protein putative aromatic gene for a BLi03991 3.29 4.33 3.86 13.22 12.18 compounds specific putative dioxygenase protein putative decarboxylase/ gene for a BLi03993 3.84 5.22 10.49 22.93 24.21 dehydratase putative protein close homolog to DhaS gene for a BLi03994 5.33 9.09 10.32 22.37 19.12 aldehyde hypothetical dehydrogenase protein putative transcriptional gene for a BLi03995 1.82 5.01 7.61 12.47 15.76 regulator putative protein unknown function gene for a BLi03996 1.52 2.34 4.24 8.76 10.13 conserved hypothetical protein putative phage protein gene for a BLi01482 1.07 2.69 2.55 3.27 3.19 putative protein putative phage protein gene for a BLi01480 1.26 2.64 2.69 3.60 3.37 putative protein putative phage capsid gene for a BLi01470 3.21 14.00 9.43 9.07 5.59 protein putative protein putative phage protein gene for a BLi01466 2.04 2.57 3.21 6.12 5.04 putative protein putative portal protein gene for a BLi01465 1.85 3.40 3.52 6.43 5.22 putative protein unknown function ykoP BLi03288 1.66 3.60 2.04 3.67 1.90 metalloregulation DNA- mrgA BLi03480 3.03 3.23 2.60 1.62 1.57 binding stress protein unknown function - yvtA BLi03481 9.99 3.41 1.30 1.61 1.00 similar to HtrA-like serine protease unknown function - yvqH BLi03496 0.96 3.55 9.12 12.94 22.11 similar to unknown function proteins from B. subtilis unknown function yvqI BLi03497 0.96 2.79 6.83 10.39 21.62 unknown function - ycbA BLi00275 0.90 1.43 2.31 3.81 5.46 similar to two- component sensor histidine kinase [YcbB] transcriptional repressor kdgR BLi03828 0.77 1.99 2.15 4.81 8.02 of the pectin utilization operon unknown function - ydhQ BLi02559 1.02 1.31 1.56 3.12 3.48 similar to transcriptional regulator (GntR family) unknown function - ycnI BLi00479 3.74 5.12 3.84 8.23 3.18 similar to unknown function proteins unknown function - ycnJ BLi00480 4.04 5.28 4.59 10.74 3.79 similar to copper export protein unknown function - ycnK BLi00481 4.60 10.14 8.17 14.46 8.30 similar to transcriptional regulator (DeoR family) assimilatory nitrate nasB BLi00482 18.17 22.63 20.62 25.24 16.47 reductase (electron transfer subunit) assimilatory nitrate nasC BLi00483 11.07 22.08 20.33 21.01 20.53 reductase (catalytic subunit) assimilatory nitrite nasD BLi00484 6.63 10.58 15.49 20.64 12.06 reductase (subunit) assimilatory nitrite nasE BLi00485 10.63 23.53 27.51 42.63 21.86 reductase (subunit) uroporphyrin-III C- nasF BLi00486 8.08 37.38 22.44 36.50 18.60 methyltransferase unknown function gene for a BLi00236 2.98 5.18 6.04 7.59 8.29 hypothetical protein unknown function - yckI BLi00418 3.21 3.55 3.01 3.09 2.03 similar to glutamine ABC transporter (ATP- binding protein) unknown function gene for a BLi00413 4.35 4.40 3.28 5.49 3.05 hypothetical protein phosphoenolpyruvate pckA BLi03197 0.90 5.90 7.35 28.58 10.77 carboxykinase translocation-dependent tasA BLi02637 4.15 3.31 1.71 4.69 1.35 antimicrobial spore component unknown function yqxM BLi02639 2.63 3.46 1.74 4.87 1.44 nuclease inhibitor dinB BLi02244 1.44 5.18 6.35 5.81 5.65 unknown function - yngK BLi02129 1.33 4.09 4.09 5.89 4.03 similar to unknown function proteins unknown function gene for a BLi04184 1.97 5.16 5.07 4.72 4.39 hypothetical protein unknown function gene for a BLi04185 1.24 2.90 2.25 6.77 5.26 hypothetical protein putative malate gene for a BLi04208 1.00 5.95 3.71 16.98 2.98 synthase (EC 4.1.3.2) putative protein unknown function - yvcK BLi03724 1.54 2.78 3.11 2.83 3.06 similar to unknown

function proteins ATP-dependent Clp clpP BLi03710 2.62 4.39 3.50 2.56 1.85 protease proteolytic subunit (class III heat- shock protein) unknown function ykyB BLi01615 1.09 2.14 2.89 4.52 3.35 unknown function gene for a BLi02232 1.07 1.66 4.21 4.17 3.71 hypothetical protein putative hydrolase gene for a BLi02233 1.29 3.38 8.50 12.34 6.57 putative protein unknown function - yfhF BLi00876 1.23 1.65 2.40 3.02 3.01 similar to cell-division inhibitor unknown function gene for a BLi01622 0.95 1.19 2.52 3.81 3.13 hypothetical protein putative glycerol gene for a BLi00828 2.03 5.17 6.18 5.69 6.48 dehydrogenase (EC putative 1.1.1.6) protein putative Na(+)-linked D- gene for a BLi00817 3.91 6.40 13.40 12.49 17.09 alanine glycine putative permease protein unknown function - ytcA BLi00810 1.40 3.88 4.32 4.56 4.07 similar to NDP-sugar dehydrogenase unknown function - ymcA BLi01926 1.14 2.26 2.58 2.13 3.69 similar to unknown function proteins unknown function gene for a BLi01933 6.35 5.54 0.93 3.71 0.97 conserved hypothetical protein NADH dehydrogenase gene for a BLi01934 6.56 4.40 0.95 3.87 0.98 (EC 1.6.99.3) NADH- ubiquinone oxidoreductase 51 kDa subunit (EC 1.6.5.3) unknown function - yjiC BLi01948 1.23 3.15 1.89 3.04 1.56 similar to macrolide glycosyl transferase gamma- ggt BLi01364 1.81 8.65 7.15 8.76 6.61 glutamyltranspeptidase putative beta-lactamase gene for a BLi00328 1.12 1.52 3.83 5.64 5.50 protein putative protein unknown function - ysfD BLi00329 0.95 1.18 3.09 3.14 3.84 similar to glycolate oxidase subunit unknown function ycdC BLi00343 1.20 1.78 4.31 4.81 4.79 putative transcriptional gene for a BLi00353 1.32 1.58 3.25 3.00 2.74 regulatory protein putative protein unknown function - yceC BLi00354 2.21 3.05 4.99 4.72 4.73 similar to tellurium resistance protein unknown function - yceD BLi00355 2.36 3.85 5.35 4.14 4.85 similar to tellurium resistance protein unknown function - yceE BLi00356 2.09 3.65 5.92 5.42 7.25 similar to tellurium resistance protein unknown function - yceF BLi00357 1.62 1.98 2.98 2.82 3.29 similar to tellurium resistance protein nitrate transporter nasA BLi00365 1.65 2.15 3.28 2.26 2.73 NH3-dependent NAD+ nadE BLi00370 1.51 3.29 3.25 2.75 2.47 synthetase unknown function - ycgL BLi00372 1.44 2.98 3.02 2.14 1.72 similar to unknown function proteins unknown function - ycgM BLi00373 1.17 10.73 14.51 18.05 2.61 similar to proline oxidase unknown function - ycgN BLi00374 1.20 7.65 8.25 12.14 1.88 similar to 1-pyrroline-5- carboxylate dehydrogenase unknown function - ycgO BLi00375 1.13 2.14 3.79 3.37 1.90 similar to proline permease unknown function - yqfA BLi02729 1.33 1.75 3.91 4.88 5.84 similar to unknown function proteins close homolog to HemH gene for a BLi04115 1.44 1.91 1.67 3.47 3.20 ferrochelatase hypothetical protein unknown function gene for a BLi04116 1.66 2.25 1.84 7.40 8.31 hypothetical protein putative ABC gene for a BLi04117 2.08 2.88 2.08 9.26 10.49 transporter ATP-binding putative protein protein unknown function gene for a BLi04118 1.08 1.72 2.23 3.90 6.80 hypothetical protein unknown function gene for a BLi04119 5.53 1.72 1.42 5.46 4.59 hypothetical protein unknown function gene for a BLi04120 1.68 2.29 2.02 7.63 7.32 conserved hypothetical protein unknown function gene for a BLi04124 1.19 2.46 3.94 4.59 6.77 conserved hypothetical protein putative bacteriocin gene for a BLi04126 1.59 3.71 4.73 6.92 6.89 formation protein putative protein putative bacteriocin gene for a BLi04128 1.49 2.73 3.80 4.16 6.01 formation protein putative protein ABC transporter cydD BLi04131 1.17 1.89 2.07 3.27 3.26 required for expression of cytochrome bd (ATP- binding protein) ABC transporter cydC BLi04132 1.19 2.33 1.80 4.73 3.59 required for expression of cytochrome bd (ATP- binding protein) cytochrome bd cydB BLi04133 0.90 1.98 1.71 3.50 3.09 ubiquinol oxidase (subunit II) cytochrome bd cydA BLi04134 0.92 2.44 2.15 4.48 3.32 ubiquinol oxidase (subunit I) family serine protease, clpP BLi03615 2.62 4.39 3.50 2.56 1.85 possible phage related unknown function - yciC BLi00765 9.94 5.08 0.41 9.70 0.44 similar to unknown function proteins citrate synthase I citA BLi01010 1.10 1.81 3.51 5.78 8.09 (minor) unknown function - yueE BLi03372 1.27 4.43 3.09 4.19 4.28 similar to unknown function proteins threonyl-tRNA thrZ BLi00234 2.34 4.75 4.27 5.60 5.26 synthetase (minor) ribose ABC transporter rbsA BLi03843 1.28 4.95 1.81 2.27 2.60 (ATP-binding protein) ribose ABC transporter rbsB BLi03845 2.09 5.61 2.33 4.47 2.79 (ribose-binding protein) alpha-acetolactate alsD BLi03847 5.84 22.15 7.42 38.12 9.31 decarboxylase alpha-acetolactate alsS BLi03848 5.86 44.43 - 40.12 10.29 synthase 17.73 76.83 9.41 143.53 23.96 transcriptional regulator alsR BLi03849 1.75 8.78 7.50 8.75 4.33 of the alpha- acetolactate operon unknown function - ywrD BLi03850 2.28 5.04 3.15 4.71 3.15 similar to gamma- glutamyl transferase unknown function - ywrC BLi03851 2.16 5.23 3.08 7.25 3.90 similar to transcriptional regulator (Lrp/AsnC family) unknown function - ywrB BLi03852 1.37 3.18 2.18 3.40 2.41 similar to chromate transport protein unknown function - ywqE BLi03854 1.45 3.05 2.83 3.77 2.59 similar to capsular polysaccharide biosynthesis unknown function - ywqC BLi03855 1.44 3.43 2.96 4.29 2.82 similar to capsular polysaccharide biosynthesis unknown function gene for a BLi03108 1.72 3.11 1.60 6.93 2.50 conserved hypothetical protein putative transcriptional gene for a BLi03548 1.15 3.37 3.93 5.48 3.52 regulator putative protein unknown function - yabE BLi00053 0.83 1.43 1.70 3.29 3.20 similar to cell wall- binding protein modulator of CtsR mcsB BLi00103 1.84 3.21 3.40 2.32 1.97 repression class III stress clpC BLi00104 2.29 5.57 4.44 2.80 2.49 response-related ATPase RNA polymerase sigH BLi00116 1.52 2.67 2.74 2.97 3.04 sigma-H factor (sigma- 30) putative DNA-binding gene for a BLi04317 1.67 3.09 4.94 2.87 2.53 protein putative protein putative am gene for a BLi01175 6.20 19.95 19.06 20.96 18.99 putative protein asparagine synthetase asnO BLi01174 2.16 5.61 5.19 5.46 5.89 signal peptidase I sipS BLi00675 1.24 2.55 2.48 3.30 3.28 unknown function yeaA BLi00672 1.21 3.04 3.59 5.20 4.83 unknown function gene for a BLi00669 1.16 2.35 3.72 6.32 4.27 hypothetical protein unknown function gene for a BLi00668 1.22 3.68 4.32 7.18 4.47 hypothetical protein unknown function - ywaE BLi00667 1.18 2.16 2.87 3.47 4.12 similar to transcriptional regulator (MarR family) unknown function - yvdH BLi00660 1.13 1.96 2.31 3.45 3.27 similar to maltodextrin transport system permease unknown function - ycdH BLi03213 21.12 7.72 1.98 4.50 1.00 similar to ABC transporter (binding protein) anthranilate synthase trpE BLi02403 1.96 26.10 22.29 19.75 23.14 anthranilate trpD BLi02402 1.50 10.22 13.01 11.88 13.42 phosphoribosyl transferase indol-3-glycerol trpC BLi02401 1.50 12.08 14.43 11.15 15.42 phosphate synthase phosphoribosyl trpF BLi02400 1.43 6.15 12.79 11.61 12.06 anthranilate isomerase tryptophan synthase trpB BLi02399 1.50 9.20 13.19 11.39 14.53 (beta subunit) tryptophan synthase trpA BLi02398 1.03 5.97 7.01 5.26 7.49 (alpha subunit) histidinol-phosphate hisC BLi02397 1.56 6.27 5.16 5.05 5.96 amino transferase/ tyrosine and phenylalanine amino transferase prephenate tyrA BLi02396 1.07 2.31 4.17 4.00 3.06 dehydrogenase 5- aroE BLi02395 1.14 3.90 4.02 3.61 3.89 enolpyruvoylshikimate- 3-phosphate synthase unknown function - ypiB BLi02393 1.69 5.02 5.60 6.15 6.43 similar to unknown function proteins unknown function - yqeH BLi02760 1.01 2.79 2.99 3.56 3.55 similar to unknown function proteins unknown function yqeF BLi02763 1.08 2.86 3.45 3.56 3.28 two-component dctR BLi02781 1.17 4.50 4.05 4.44 3.82 response regulator involved in C4- dicarboxylate transport putative two-component gene for a BLi02787 1.19 5.14 2.62 4.67 2.86 response regulator putative involved in degradative protein

enzyme and competence regulation 6-phospho-3- hxlB BLi02804 1.25 3.15 1.35 2.56 2.48 hexuloisomerase 3-hexulose-6-phosphate hxlA BLi02805 2.56 4.13 2.48 3.73 2.10 synthase unknown function yppF BLi02362 1.15 3.31 2.69 4.07 8.62 alkyl hydroperoxide ahpF BLi04292 2.30 3.07 2.33 2.55 2.10 reductase (large subunit)/NADH dehydrogenase alkyl hydroperoxide ahpC BLi04291 3.17 3.21 2.35 2.40 1.50 reductase (small subunit) putative gene for a BLi04251 1.09 2.25 2.06 3.02 2.97 methylmalonate- putative semialdehyde protein dehydrogenase [acylating] (EC 1.2.1.27) unknown function - ykoU BLi01494 1.91 5.27 5.01 5.64 6.04 similar to ATP- dependent DNA ligase unknown function - ykoM BLi01493 2.13 5.84 6.31 7.24 5.83 similar to transcriptional regulator (MarR family) unknown function - ydfS BLi01489 2.03 6.03 4.84 13.04 6.72 similar to unknown function proteins unknown function gene for a BLi01488 7.88 73.93 103.66 146.73 54.19 hypothetical protein unknown function - yrkC BLi01487 3.01 38.73 35.62 62.29 20.60 similar to unknown function proteins putative glycosyl gene for a BLi01486 2.19 8.60 10.08 16.48 13.52 hydrolase/lysozyme putative protein unknown function yncE BLi03150 1.84 5.91 7.32 11.97 10.11 unknown function - yhbD BLi00958 1.45 5.23 7.84 17.05 12.08 similar to unknown function proteins from B. subtilis unknown function - yvlB BLi03752 1.45 3.88 7.43 10.22 12.00 similar to unknown function proteins unknown function - yvlD BLi03750 0.93 1.31 2.96 4.00 4.49 similar to unknown function proteins unknown function - yxkH BLi04151 0.77 1.67 4.79 5.99 5.82 similar to unknown function proteins transcriptional regulator pucR BLi03436 1.91 2.82 4.27 4.96 4.20 of puc genes transcriptional activator gltC BLi02163 1.15 3.54 4.43 5.21 5.34 of the glutamate synthase operon unknown function gene for a BLi03570 2.58 3.81 6.18 5.44 5.03 hypothetical protein putative amidase gene for a BLi03571 1.95 2.40 2.11 3.45 2.96 putative protein unknown function - ylbP BLi01727 1.56 4.57 3.28 3.27 2.35 similar to unknown function proteins putative ABC gene for a BLi03212 8.10 4.42 1.28 4.52 1.45 transporter/amino acid putative permease protein putative nitrogen gene for a BLi01176 8.33 30.73 31.25 33.74 30.23 regulatory protein P-II putative protein unknown function gene for a BLi00719 6.47 17.69 14.11 18.09 17.23 hypothetical protein unknown function - ywpF BLi03871 1.51 3.86 3.82 5.12 3.69 similar to unknown function proteins unknown function yvlA BLi03753 1.32 4.63 8.68 10.55 11.65 unknown function - ywbD BLi04060 2.49 2.17 3.88 6.12 4.20 similar to unknown function proteins unknown function gene for a BLi00654 1.06 6.05 3.60 4.45 3.81 conserved hypothetical protein putative phage tail gene for a BLi01476 1.01 4.36 2.24 3.31 2.37 protein putative protein peptidyl methionine msrA BLi02303 1.83 4.46 3.66 3.71 2.74 sulfoxide reductase unknown function gene for a BLi00235 5.03 10.82 7.36 11.39 9.29 hypothetical protein

Example 3

Genes that are Markedly Induced Specifically Under Nitrogen Deficiency

[0233]As is shown in Table 1, a total of 67 of all the Bacillus licheniformis DSM13 genes investigated in Example 1 under the conditions of nitrogen deficiency were induced by at least a factor of 8 at any of the measured times. Among these were also 18 genes that were also induced by the deficiency of at least one other compound (data not shown) and which therefore cannot be regarded as specific signals for a nitrogen deficiency.

[0234]Consequently there remain 49 genes that were induced by at least a factor 8 at any of the measured times and may be classified as comparatively specific. They are compiled in Table 2 below. The column headers are the same as in the preceding example. In addition, the first column indicates the sequence numbers of the particular DNA and amino acid sequences in the sequence listing of the present application. Specific features of the particular sequences, which appear as free text in the sequence listing have been added under the heading Gene name/gene function.

TABLE-US-00002 TABLE 2 The 62 Bacillus licheniformis DSM13 genes determined in Example 1, whose specific induction (by at least a factor of 8) under nitrogen deficiency was observed at any of the measured times (explanations: see text). SEQ Protein name/ ID Gene function/ NO. Notes on the gene Gene BLi--Nr Transition 30 min 60 min 90 min 120 min 1, 2 unknown function - yqjN BLi02550 1.56 2.68 4.48 8.50 8.65 similar to amino acid degradation The first codon is translated as methionine. 3, 4 nitrogen-regulated nrgB BLi03889 3.61 6.92 5.04 9.99 6.67 PII-like protein 5, 6 ammonium nrgA BLi03888 3.04 5.71 3.91 9.56 7.31 transporter 7, 8 serine protease Do htrA BLi01390 7.91 8.10 2.03 1.93 1.67 (heat-shock protein) 9, 10 putative serine gene for a BLi00301 2.31 13.40 7.51 15.91 11.55 protease putative The first codon is protein translated as methionine. 11, transcriptional glnR BLi01992 4.38 11.07 7.61 9.77 8.25 12 repressor of the glutamine synthetase gene 13, glutamine glnA BLi01993 5.69 9.51 8.60 9.06 8.61 14 synthetase 15, close homolog to gene for a BLi03994 5.33 9.09 10.32 22.37 19.12 16 DhaS aldehyde hypothetical dehydrogenase protein 17, putative gene for a BLi03995 1.82 5.01 7.61 12.47 15.76 18 transcriptional putative regulator protein 19, unknown function gene for a BLi03996 1.52 2.34 4.24 8.76 10.13 20 The first codon is conserved translated as hypothetical methionine. protein 21, putative phage gene for a BLi01470 3.21 14.00 9.43 9.07 5.59 22 capsid protein putative protein 23, unknown function - yvtA BLi03481 9.99 3.41 1.30 1.61 1.00 24 similar to HtrA-like serine protease 25, transcriptional kdgR BLi03828 0.77 1.99 2.15 4.81 8.02 26 repressor of the pectin utilization operon 27, unknown function - ycnI BLi00479 3.74 5.12 3.84 8.23 3.18 28 similar to unknown function proteins 29, unknown function - ycnJ BLi00480 4.04 5.28 4.59 10.74 3.79 30 similar to copper export protein 31, unknown function - ycnK BLi00481 4.60 10.14 8.17 14.46 8.30 32 similar to transcriptional regulator (DeoR family) 33, assimilatory nitrate nasB BLi00482 18.17 22.63 20.62 25.24 16.47 34 reductase (electron transfer subunit) 35, assimilatory nitrate nasC BLi00483 11.07 22.08 20.33 21.01 20.53 36 reductase (catalytic subunit) 37, assimilatory nitrite nasD BLi00484 6.63 10.58 15.49 20.64 12.06 38 reductase (subunit) The first codon is translated as methionine. 39, uroporphyrin-III C- nasF BLi00486 8.08 37.38 22.44 36.50 18.60 40 methyltransferase 41, unknown function gene for a BLi00236 2.98 5.18 6.04 7.59 8.29 42 hypothetical protein 43, phosphoenolpyruvate pckA BLi03197 0.90 5.90 7.35 28.58 10.77 44 carboxykinase 45, putative malate gene for a BLi04208 1.00 5.95 3.71 16.98 2.98 46 synthase (EC putative 4.1.3.2) protein 47, putative hydrolase gene for a BLi02233 1.29 3.38 8.50 12.34 6.57 48 putative protein 49, putative Na(+)- gene for a BLi00817 3.91 6.40 13.40 12.49 17.09 50 linked D-alanine putative glycine permease protein 51, gamma- ggt BLi01364 1.81 8.65 7.15 8.76 6.61 52 glutamyltranspeptidase 53, unknown function gene for a BLi04116 1.66 2.25 1.84 7.40 8.31 54 hypothetical protein 55, putative ABC gene for a BLi04117 2.08 2.88 2.08 9.26 10.49 56 transporter ATP- putative binding protein protein 57, unknown function - yciC BLi00765 9.94 5.08 0.41 9.70 0.44 58 similar to unknown function proteins 59, citrate synthase I citA BLi01010 1.10 1.81 3.51 5.78 8.09 60 (minor) 61, transcriptional alsR BLi03849 1.75 8.78 7.50 8.75 4.33 62 regulator of the alpha-acetolactate operon 63, putative ammonium gene for a BLi01175 6.20 19.95 19.06 20.96 18.99 64 transporter putative protein 65, unknown function - ycdH BLi03213 21.12 7.72 1.98 4.50 1.00 66 similar to ABC transporter (binding protein) 67, anthranilate trpE BLi02403 1.96 26.10 22.29 19.75 23.14 68 synthase The first codon is translated as methionine. 69, anthranilate trpD BLi02402 1.50 10.22 13.01 11.88 13.42 70 phosphoribosyl transferase 71, indol-3-glycerol trpC BLi02401 1.50 12.08 14.43 11.15 15.42 72 phosphate synthase 73, phosphoribosyl trpF BLi02400 1.43 6.15 12.79 11.61 12.06 74 anthranilate isomerase 75, tryptophan synthase trpB BLi02399 1.50 9.20 13.19 11.39 14.53 76 (beta subunit)) 77, unknown function yppF BLi02362 1.15 3.31 2.69 4.07 8.62 78 79, unknown function - ydfS BLi01489 2.03 6.03 4.84 13.04 6.72 80 similar to unknown function proteins 81, unknown function gene for a BLi01488 7.88 73.93 103.66 146.73 54.19 82 hypothetical protein 83, unknown function - yrkC BLi01487 3.01 38.73 35.62 62.29 20.60 84 similar to unknown function proteins 85, putative glycosyl gene for a BLi01486 2.19 8.60 10.08 16.48 13.52 86 hydrolase/ putative lysozyme protein 87, unknown function yncE BLi03150 1.84 5.91 7.32 11.97 10.11 88 89, unknown function - yvlB BLi03752 1.45 3.88 7.43 10.22 12.00 90 similar to unknown function proteins 91, putative ABC gene for a BLi03212 8.10 4.42 1.28 4.52 1.45 92 transporter/amino putative acid permease protein 93, putative nitrogen gene for a BLi01176 8.33 30.73 31.25 33.74 30.23 94 regulatory protein putative P-II protein The first codon is translated as methionine. 95, unknown function gene for a BLi00719 6.47 17.69 14.11 18.09 17.23 96 The first codon is hypothetical translated as protein methionine. 97, unknown function yvlA BLi03753 1.32 4.63 8.68 10.55 11.65 98

[0235]In Table 3 below are listed the same 49 genes once more in the order of observed strength of induction. The corresponding terms in English are indicated in this order in the description section. The arrangement of the subject matter of the claims follows the reverse order.

TABLE-US-00003 TABLE 3 The 49 Bacillus licheniformis DSM13 genes determined in Example 1, whose specific induction (by at least a factor of 8) under nitrogen deficiency was observed at any of the measured times, are listed in descending order of the maximum value (last column) measured in each case. SEQ ID Protein name/Gene function/ NO. Notes on the gene Gene BLi-Nr max. 81, unknown function gene for a BLi01488 146.73 82 hypothetical protein 83, unknown function - similar to yrkC BLi01487 62.29 84 unknown function proteins 39, uroporphyrin-III C-methyltransferase nasF BLi00486 37.38 40 93, putative nitrogen regulatory protein P- gene for a putative BLi01176 33.74 94 II protein The first codon is translated as methionine. 43, phosphoenolpyruvate carboxykinase pckA BLi03197 28.58 44 67, anthranilate synthase trpE BLi02403 26.10 68 The first codon is translated as methionine. 33, assimilatory nitrate reductase nasB BLi00482 25.24 34 (electron transfer subunit) 15, close homolog to DhaS aldehyde gene for a BLi03994 22.37 16 dehydrogenase hypothetical protein 35, assimilatory nitrate reductase nasC BLi00483 22.08 36 (catalytic subunit) 65, unknown function - similar to ABC ycdH BLi03213 21.12 66 transporter (binding protein) 63, putative ammonium transporter gene for a putative BLi01175 20.96 64 protein 37, assimilatory nitrite reductase nasD BLi00484 20.64 38 (subunit) The first codon is translated as methionine. 95, unknown function gene for a BLi00719 18.09 96 The first codon is translated as hypothetical protein methionine. 49, putative Na(+)-linked D-alanine gene for a putative BLi00817 17.09 50 glycine permease protein 45, putative malate synthase (EC 4.1.3.2) gene for a putative BLi04208 16.98 46 protein 85, putative glycosyl hydrolase/ gene for a putative BLi01486 16.48 86 lysozyme protein 9, putative serine protease gene for a putative BLi00301 15.91 10 The first codon is translated as protein methionine. 17, putative transcriptional regulator gene for a putative BLi03995 15.76 18 protein 71, indol-3-glycerol phosphate synthase trpC BLi02401 15.42 72 75, tryptophan synthase (beta subunit) trpB BLi02399 14.53 76 31, unknown function - similar to ycnK BLi00481 14.46 32 transcriptional regulator (DeoR family) 21, putative phage capsid protein gene for a putative BLi01470 14.00 22 protein 69, anthranilate phosphoribosyl trpD BLi02402 13.42 70 transferase 79, unknown function - similar to ydfS BLi01489 13.04 80 unknown function proteins 73, phosphoribosyl anthranilate trpF BLi02400 12.79 74 isomerase 47, putative hydrolase gene for a putative BLi02233 12.34 48 protein 89, unknown function - similar to yvlB BLi03752 12.00 90 unknown function proteins 87, unknown function yncE BLi03150 11.97 88 97, unknown function yvlA BLi03753 11.65 98 11, transcriptional repressor of the glnR BLi01992 11.07 12 glutamine synthetase gene 29, unknown function - similar to copper ycnJ BLi00480 10.74 30 export protein 55, putative ABC transporter ATP-binding gene for a putative BLi04117 10.49 56 protein protein 19, unknown function gene for a BLi03996 10.13 20 The first codon is translated as conserved methionine. hypothetical protein 3, 4 nitrogen-regulated PII-like protein nrgB BLi03889 9.99 23, unknown function - similar to HtrA- yvtA BLi03481 9.99 24 like serine protease 57, unknown function - similar to yciC BLi00765 9.94 58 unknown function proteins 5, 6 ammonium transporter nrgA BLi03888 9.56 13, glutamine synthetase glnA BLi01993 9.51 14 61, transcriptional regulator of the alpha- alsR BLi03849 8.78 62 acetolactate operon 51, gamma-glutamyltranspeptidase ggt BLi01364 8.76 52 1, 2 unknown function - similar to amino yqjN BLi02550 8.65 acid degradation The first codon is translated as methionine. 77, unknown function yppF BLi02362 8.62 78 53, unknown function gene for a BLi04116 8.31 54 hypothetical protein 41, unknown function gene for a BLi00236 8.29 42 hypothetical protein 27, unknown function - similar to ycnI BLi00479 8.23 28 unknown function proteins 7, 8 serine protease Do (heat-shock htrA BLi01390 8.10 protein) 91, putative ABC transporter/amino acid gene for a putative BLi03212 8.10 92 permease protein 59, citrate synthase I (minor) citA BLi01010 8.09 60 25, transcriptional repressor of the pectin kdgR BLi03828 8.02 26 utilization operon

Example 4

Real Time RT-PCR Quantification of Selected Genes

[0236]Real time RT-PCR enables the absolute numbers of molecules of specific transcripts in a sample to be determined. The LightCycler (Roche Diagnostics, Penzberg, Germany) was utilized in combination with the "SYBR Green I" kit (Roche Diagnostics). This method is based on detecting the binding of the fluorescent dye to double-stranded DNA. The specific mRNA molecules were quantified as described in Tobisch et al. (2003; Quantification of Bacterial mRNA by One-Step RT-PCR Using the LightCycler System; BIOCHEMICA, Volume 3, pages 5 to 8), with an external standard curve being established for each mRNA to be studied.

[0237]For this purpose, dilutions of known concentrations of in vitro transcripts of the mRNAs determined in Example 1 and listed in Example 2 were measured with the aid of the LightCycler, and then the standard curve was generated using the apparatus-specific software. For this purpose, primers, which are specific for each mRNA to be studied, and which enable in vitro transcripts to be synthesized and PCR amplification in the LightCycler had to be selected beforehand (with the aid of the "Array Designer" software; available from PREMIER Biosoft International, Palo Alto, USA).

[0238]After generating the external standard curve, measurement in the LightCycler was begun. Two different dilutions of each sample to be analyzed were used for the measurements. The particular primers were then used in the LightCycler run to amplify the specific transcript, and the incorporation of the dye was then measured.

[0239]Using the LightCycler software and the script accessible on the website http://molbiol.ru/ger/scripts/01_--07.html (12.2.2004), it was possible to determine the exact number of molecules for selected transcripts. The values obtained in this manner and their relations to one another were then used to confirm the induction values indicated in Tables 1 and 2 for the selected transcripts.

Example 5

Real Time RT-PCR Quantification of the Genes glnA, nrgB and tnrA

[0240]The genes glnA (SEQ ID NO. 13), nrgB (SEQ ID NO. 3) and tnrA were subjected to an additional Real-Time-RT-PCR, as described in the previous example. tnrA concerns the transcriptional regulator involved in global nitrogen regulation gene illustrated in SEQ ID NO. 99 (Data in the sequence listing: <223> tnrA--transcriptional pleiotropic regulator involved in global nitrogen regulation--BLi 01490). This gene incorporating 333 by is obtainable from B. licheniformis DSM 13 and is recorded in the abovementioned databank under the number BLi 01490. The derived amino acid sequence is listed under SEQ ID NO. 100.

[0241]The measurements were again carried out with the LightCycler instrument using the SYBR Green detection kit. The gene-specific primers were derived with the Programm Array Designer 2.0 (PREMIER Biosoft International, Palo Alto, USA) and ordered from Invitrogen (Karlsruhe, Germany). For the quantification of the specific mRNA, an external standard curve was generated with the help of an in-vitro transcript. The in-vitro transcript was constructed using a T7 polymerase, the gene-specific PCR product and the DIG-RNA-Labeling-Kit (Roche Diagnostics, Penzberg, Germany). The concentration and quality of the in-vitro transcript were determined photometrically.

[0242]A dilution series (1 ng to 100 fg final concentration) of the in-vitro transcript was prepared and mixed with MS2-RNA (final concentration 0.5 μg/μl; Roche Diagnostics) the forward and reverse primers and the LightCycler-RNA-Master-SYBR-Green I-Kit (Roche Diagnostics) as the mastermix, according to the manufacturer's instructions. The standard curve for the specific gene was generated by means of the manufacturer's notes for the RNA-Master-SYBRGreen I-Kit (amplification, segment 2, temperature: 57° C.; amplification, segment 3, incubation time: 16 s). The result of the LightCycler run was evaluated with the help of the LightCycler-Software and the Second-derivative-maximum Method, and the quality of the RT-PCR products was checked by the melting point determination.

[0243]For the measurements of the RNA samples, dilution steps from 500 ng to 5 ng of the total RNA were prepared and mixed with MS2-RNA (final concentration 0.5 μg/μl), the specific primer pairs and the LightCycler-RNA-Master-SYBR-Green I-Kit. The same conditions that were also used with the LightCycler instrument for generating the standard curve were used for the Real-time-RT-PCR run.

[0244]The negative control was RNase-free water with MS2-RNA in a final concentration of 0.5 μg/μl. A selected dilution of a sample from the standard curve was carried out in each case during the LightCycler run.

[0245]The LightCycler measurements were evaluated with the LightCycler-specific software. The molecular quantities of the specific RNA in the measured sample were determined with the help of the standard curve and the Second derivative maximum method.

[0246]The results compiled in the Table 4 below refer in each case to the molecular quantities during the logarithmic phase, that in each case were set to 1 (control). Samples were taken at the time of onset of the nitrogen limitation that is linked, as described above, with the transition into the stationary phase (transient phase), as well as 30, 60 and 120 minutes after this transition.

TABLE-US-00004 TABLE 4 Real-Time-RT-PCR-Quantification of the Genes glnA, nrgB and tnrA with the onset of a nitrogen deficiency and the consequent transition into the stationary phase glnA nrgB tnrA Control 1 1 1 Transient Phase 11.5 15 10.65 30 min 12.5 14.25 15.24 stationary 60 min 5.5 5.75 9,375 stationary 120 min 3.3 4,875 8.85 stationary

[0247]It can be seen that in this measurement, all three genes glnA, nrgB and tnrA are strongly expressed at the onset of a nitrogen deficiency and the transition into the stationary phase linked to this, in the case of tnrA are expressed up to 15.24 fold greater than during the exponential growth phase and the associated optimal nitrogen supply. Consequently, they are suitable inventive marker genes for indicating an onset of nitrogen deficiency.

DESCRIPTION OF THE FIGURES

[0248]FIG. 1: Schematic illustration of the At-line monitoring of a bioprocess with the inventive electrical DNA chips

The real-time monitoring of the bioprocess advantageously involves the following steps: [0249]1. Sampling, for example from the fermentation of a microorganism; [0250]2. Cell disruption by routine methods; [0251]3. RNA isolation by routine methods; [0252]4. Hybridization on a chip inventively loaded with nucleic acids (for example DNA) or nucleic acid analogs (for example difficultly hydrolyzable, analogously structured compounds); [0253]5. Acquisition of the electrical signals from a suitably constructed electrochip; alternatively, the recording of optical signals from an optical DNA chip would also be possible; [0254]6. Preferably computer-supported data analysis.When electrical chips are used, an approximate total analysis time is less than 2 hours under the present state of development; with conventional optical DNA chips it takes about 12 hours.

Sequence CWU 1

10011605DNABacillus licheniformis DSM 13gene(1)..(1605)yqjN - unknown function - similar to amino acid degradation - BLi02550 1gtg aag tgg cag aca gtg gaa gaa ctg aaa cgg ctg ctc att tct ctt 48Val Lys Trp Gln Thr Val Glu Glu Leu Lys Arg Leu Leu Ile Ser Leu1 5 10 15gtt caa tac gaa agc atc tca ggc acg gcc gga gaa gtg gcg ctt gcg 96Val Gln Tyr Glu Ser Ile Ser Gly Thr Ala Gly Glu Val Ala Leu Ala 20 25 30aaa tat atg cat gat gtc tta aag gac cgt tcg tac ttt caa aag aat 144Lys Tyr Met His Asp Val Leu Lys Asp Arg Ser Tyr Phe Gln Lys Asn 35 40 45cct gaa tac tta aag ctt cac ccg atg gag gat ggg cgc tat ttt tta 192Pro Glu Tyr Leu Lys Leu His Pro Met Glu Asp Gly Arg Tyr Phe Leu 50 55 60acc gcc ctc gtc aaa aag gaa aag aaa tca aat acg gtg ctc ctc ttg 240Thr Ala Leu Val Lys Lys Glu Lys Lys Ser Asn Thr Val Leu Leu Leu65 70 75 80agc cat ttt gac gtg gtc gat acg gca gac tac gga gaa ttc aaa cat 288Ser His Phe Asp Val Val Asp Thr Ala Asp Tyr Gly Glu Phe Lys His 85 90 95atg gca tgc aaa gtc ccg gaa ctc atg gac cta tta aat gat aaa aaa 336Met Ala Cys Lys Val Pro Glu Leu Met Asp Leu Leu Asn Asp Lys Lys 100 105 110gaa ctt ttg cct gaa cgg gtc aaa aaa gac ctg gaa agc ggc gaa tgg 384Glu Leu Leu Pro Glu Arg Val Lys Lys Asp Leu Glu Ser Gly Glu Trp 115 120 125ctg ttc ggt agg gga agc atg gat atg aaa gcg ggt ctt gct gtt caa 432Leu Phe Gly Arg Gly Ser Met Asp Met Lys Ala Gly Leu Ala Val Gln 130 135 140ttt tct atg ctg gaa cgg gcg atg aac gga gaa ttt gac gga aac ctt 480Phe Ser Met Leu Glu Arg Ala Met Asn Gly Glu Phe Asp Gly Asn Leu145 150 155 160cta tta att acg gtt ccg gac gaa gaa gtc aac tcg cag ggg atg ctt 528Leu Leu Ile Thr Val Pro Asp Glu Glu Val Asn Ser Gln Gly Met Leu 165 170 175gaa gca gta cct gta ctg aaa gct tta gaa aaa gaa ttt ggc ctt cat 576Glu Ala Val Pro Val Leu Lys Ala Leu Glu Lys Glu Phe Gly Leu His 180 185 190tat aca gcg tgc tta aac tgt gag ccg atg ttt gaa aaa cat ccg ggt 624Tyr Thr Ala Cys Leu Asn Cys Glu Pro Met Phe Glu Lys His Pro Gly 195 200 205gat ccg aag cag tac gtc tat tca ggc agc atc ggc aaa gcg ctt gcc 672Asp Pro Lys Gln Tyr Val Tyr Ser Gly Ser Ile Gly Lys Ala Leu Ala 210 215 220gga ttt ttc tgc aaa ggg gtt gag acg cat gtc ggc gag ccg ttt gca 720Gly Phe Phe Cys Lys Gly Val Glu Thr His Val Gly Glu Pro Phe Ala225 230 235 240ggg gtg aat gcg aat cta atg gtg tct gag atc aac agg ctg ctc gaa 768Gly Val Asn Ala Asn Leu Met Val Ser Glu Ile Asn Arg Leu Leu Glu 245 250 255tta aac gtc gac tat tgc gaa gaa gcg gac gga gag gtg acg cct ccc 816Leu Asn Val Asp Tyr Cys Glu Glu Ala Asp Gly Glu Val Thr Pro Pro 260 265 270ccg gcc aat ctg atg caa aaa gat ctg aag gag gcc tac tcg gtt cag 864Pro Ala Asn Leu Met Gln Lys Asp Leu Lys Glu Ala Tyr Ser Val Gln 275 280 285act cct cac aca gcg gtg tcg ctc ttt aat gtg ctg atg atg cag cgt 912Thr Pro His Thr Ala Val Ser Leu Phe Asn Val Leu Met Met Gln Arg 290 295 300tcc gcc gcc gag ctc cat cgc atg ctg ctc gac aca gcc agg gaa gcg 960Ser Ala Ala Glu Leu His Arg Met Leu Leu Asp Thr Ala Arg Glu Ala305 310 315 320gcc gcc aat att caa aca aat atc gaa cgt aaa acg gcc gaa tat cag 1008Ala Ala Asn Ile Gln Thr Asn Ile Glu Arg Lys Thr Ala Glu Tyr Gln 325 330 335cgg ttt gaa gcc ttc acg cca att aac ccg gcc gtt tcc gtg atg acg 1056Arg Phe Glu Ala Phe Thr Pro Ile Asn Pro Ala Val Ser Val Met Thr 340 345 350ttt gag gag ctg tgg aac aaa gca cag gaa aaa gcc ggg aag ccc gag 1104Phe Glu Glu Leu Trp Asn Lys Ala Gln Glu Lys Ala Gly Lys Pro Glu 355 360 365gcg gaa cgc atc ctc aac ttt gca ttt gcg aac agg ggg gag ctt gga 1152Ala Glu Arg Ile Leu Asn Phe Ala Phe Ala Asn Arg Gly Glu Leu Gly 370 375 380gac cgc gac ttt tca acg aaa atc gtc tct gaa ttg gct tca ctc tgc 1200Asp Arg Asp Phe Ser Thr Lys Ile Val Ser Glu Leu Ala Ser Leu Cys385 390 395 400aaa gaa gat gct ccg atg atc gtt ctg ttt tac agt ccg ccg ttt tat 1248Lys Glu Asp Ala Pro Met Ile Val Leu Phe Tyr Ser Pro Pro Phe Tyr 405 410 415ccg gct gtt tct tca aag gac gat cct ctg atc cga aag gtg atg aga 1296Pro Ala Val Ser Ser Lys Asp Asp Pro Leu Ile Arg Lys Val Met Arg 420 425 430gaa gtg aag cag tat gcg gcc gac agg cac gga att gaa ttt aca gaa 1344Glu Val Lys Gln Tyr Ala Ala Asp Arg His Gly Ile Glu Phe Thr Glu 435 440 445gtc caa tat ttc ccc ggc ctg tcc gat tta agc tac ctg cag ctg caa 1392Val Gln Tyr Phe Pro Gly Leu Ser Asp Leu Ser Tyr Leu Gln Leu Gln 450 455 460aaa cag tca gtc gat gcg ttc aca tcc aat atg ccg ctt ttt cat cac 1440Lys Gln Ser Val Asp Ala Phe Thr Ser Asn Met Pro Leu Phe His His465 470 475 480ggc tat tca ctt cca aaa gga cgc gaa aac gca ctg ttt gtg ccg gtc 1488Gly Tyr Ser Leu Pro Lys Gly Arg Glu Asn Ala Leu Phe Val Pro Val 485 490 495atc aat atc ggg ccg gcc ggc aaa gat cct cac aaa tgg acg gaa cgt 1536Ile Asn Ile Gly Pro Ala Gly Lys Asp Pro His Lys Trp Thr Glu Arg 500 505 510ctc cat gtg ccg ttt tca ttt gga aag ctg cca gat atc ctc cgg ctc 1584Leu His Val Pro Phe Ser Phe Gly Lys Leu Pro Asp Ile Leu Arg Leu 515 520 525gcg ata cac aag ctt ctc taa 1605Ala Ile His Lys Leu Leu 5302534PRTBacillus licheniformis DSM 13misc_feature(1)..(1)Xaa can be Val or Met. 2Xaa Lys Trp Gln Thr Val Glu Glu Leu Lys Arg Leu Leu Ile Ser Leu1 5 10 15Val Gln Tyr Glu Ser Ile Ser Gly Thr Ala Gly Glu Val Ala Leu Ala 20 25 30Lys Tyr Met His Asp Val Leu Lys Asp Arg Ser Tyr Phe Gln Lys Asn 35 40 45Pro Glu Tyr Leu Lys Leu His Pro Met Glu Asp Gly Arg Tyr Phe Leu 50 55 60Thr Ala Leu Val Lys Lys Glu Lys Lys Ser Asn Thr Val Leu Leu Leu65 70 75 80Ser His Phe Asp Val Val Asp Thr Ala Asp Tyr Gly Glu Phe Lys His 85 90 95Met Ala Cys Lys Val Pro Glu Leu Met Asp Leu Leu Asn Asp Lys Lys 100 105 110Glu Leu Leu Pro Glu Arg Val Lys Lys Asp Leu Glu Ser Gly Glu Trp 115 120 125Leu Phe Gly Arg Gly Ser Met Asp Met Lys Ala Gly Leu Ala Val Gln 130 135 140Phe Ser Met Leu Glu Arg Ala Met Asn Gly Glu Phe Asp Gly Asn Leu145 150 155 160Leu Leu Ile Thr Val Pro Asp Glu Glu Val Asn Ser Gln Gly Met Leu 165 170 175Glu Ala Val Pro Val Leu Lys Ala Leu Glu Lys Glu Phe Gly Leu His 180 185 190Tyr Thr Ala Cys Leu Asn Cys Glu Pro Met Phe Glu Lys His Pro Gly 195 200 205Asp Pro Lys Gln Tyr Val Tyr Ser Gly Ser Ile Gly Lys Ala Leu Ala 210 215 220Gly Phe Phe Cys Lys Gly Val Glu Thr His Val Gly Glu Pro Phe Ala225 230 235 240Gly Val Asn Ala Asn Leu Met Val Ser Glu Ile Asn Arg Leu Leu Glu 245 250 255Leu Asn Val Asp Tyr Cys Glu Glu Ala Asp Gly Glu Val Thr Pro Pro 260 265 270Pro Ala Asn Leu Met Gln Lys Asp Leu Lys Glu Ala Tyr Ser Val Gln 275 280 285Thr Pro His Thr Ala Val Ser Leu Phe Asn Val Leu Met Met Gln Arg 290 295 300Ser Ala Ala Glu Leu His Arg Met Leu Leu Asp Thr Ala Arg Glu Ala305 310 315 320Ala Ala Asn Ile Gln Thr Asn Ile Glu Arg Lys Thr Ala Glu Tyr Gln 325 330 335Arg Phe Glu Ala Phe Thr Pro Ile Asn Pro Ala Val Ser Val Met Thr 340 345 350Phe Glu Glu Leu Trp Asn Lys Ala Gln Glu Lys Ala Gly Lys Pro Glu 355 360 365Ala Glu Arg Ile Leu Asn Phe Ala Phe Ala Asn Arg Gly Glu Leu Gly 370 375 380Asp Arg Asp Phe Ser Thr Lys Ile Val Ser Glu Leu Ala Ser Leu Cys385 390 395 400Lys Glu Asp Ala Pro Met Ile Val Leu Phe Tyr Ser Pro Pro Phe Tyr 405 410 415Pro Ala Val Ser Ser Lys Asp Asp Pro Leu Ile Arg Lys Val Met Arg 420 425 430Glu Val Lys Gln Tyr Ala Ala Asp Arg His Gly Ile Glu Phe Thr Glu 435 440 445Val Gln Tyr Phe Pro Gly Leu Ser Asp Leu Ser Tyr Leu Gln Leu Gln 450 455 460Lys Gln Ser Val Asp Ala Phe Thr Ser Asn Met Pro Leu Phe His His465 470 475 480Gly Tyr Ser Leu Pro Lys Gly Arg Glu Asn Ala Leu Phe Val Pro Val 485 490 495Ile Asn Ile Gly Pro Ala Gly Lys Asp Pro His Lys Trp Thr Glu Arg 500 505 510Leu His Val Pro Phe Ser Phe Gly Lys Leu Pro Asp Ile Leu Arg Leu 515 520 525Ala Ile His Lys Leu Leu 5303351DNABacillus licheniformis DSM 13gene(1)..(351)nrgB - nitrogen-regulated PII-like protein - BLi03889 3atg agc ggt caa atg tat aaa gtg gaa att gta acc cgt cct aca aac 48Met Ser Gly Gln Met Tyr Lys Val Glu Ile Val Thr Arg Pro Thr Asn1 5 10 15ttt gaa aag ctg aaa acg gag ctg gcc aaa atc ggg gta acg tcc att 96Phe Glu Lys Leu Lys Thr Glu Leu Ala Lys Ile Gly Val Thr Ser Ile 20 25 30acg ttt tac aat gta cac gga tgc ggc ctg caa aaa ggg cat act gag 144Thr Phe Tyr Asn Val His Gly Cys Gly Leu Gln Lys Gly His Thr Glu 35 40 45ctt tac cgc ggg gtc aaa cgc gaa agc aat gtg tat gaa agg tta aaa 192Leu Tyr Arg Gly Val Lys Arg Glu Ser Asn Val Tyr Glu Arg Leu Lys 50 55 60gtc gaa atc gtc gtc agc aaa gtg cct gtc gaa aag gtg gcc gag acg 240Val Glu Ile Val Val Ser Lys Val Pro Val Glu Lys Val Ala Glu Thr65 70 75 80gcg caa aac gtc ctg aaa acc ggg gaa ccc ggt gac ggc aag atc ttc 288Ala Gln Asn Val Leu Lys Thr Gly Glu Pro Gly Asp Gly Lys Ile Phe 85 90 95ata tac gaa att aaa aac acc atc aac atc cgc aca ggc gag gaa ggt 336Ile Tyr Glu Ile Lys Asn Thr Ile Asn Ile Arg Thr Gly Glu Glu Gly 100 105 110cct gac gct tta taa 351Pro Asp Ala Leu 1154116PRTBacillus licheniformis DSM 13 4Met Ser Gly Gln Met Tyr Lys Val Glu Ile Val Thr Arg Pro Thr Asn1 5 10 15Phe Glu Lys Leu Lys Thr Glu Leu Ala Lys Ile Gly Val Thr Ser Ile 20 25 30Thr Phe Tyr Asn Val His Gly Cys Gly Leu Gln Lys Gly His Thr Glu 35 40 45Leu Tyr Arg Gly Val Lys Arg Glu Ser Asn Val Tyr Glu Arg Leu Lys 50 55 60Val Glu Ile Val Val Ser Lys Val Pro Val Glu Lys Val Ala Glu Thr65 70 75 80Ala Gln Asn Val Leu Lys Thr Gly Glu Pro Gly Asp Gly Lys Ile Phe 85 90 95Ile Tyr Glu Ile Lys Asn Thr Ile Asn Ile Arg Thr Gly Glu Glu Gly 100 105 110Pro Asp Ala Leu 11551212DNABacillus licheniformis DSM 13gene(1)..(1212)nrgA - ammonium transporter - BLi03888 5atg cag atg ggg gat aca gta ttt atg ttc ttt tgc gct ttg ctc gtg 48Met Gln Met Gly Asp Thr Val Phe Met Phe Phe Cys Ala Leu Leu Val1 5 10 15tgg ctg atg acg ccg ggc atc gct tta ttt tac gga ggc ctt gta agg 96Trp Leu Met Thr Pro Gly Ile Ala Leu Phe Tyr Gly Gly Leu Val Arg 20 25 30agc aaa aat gtg ctc agc acc gtt atg cac agt tta tct tca att gcc 144Ser Lys Asn Val Leu Ser Thr Val Met His Ser Leu Ser Ser Ile Ala 35 40 45atc gtt tct atc ata tgg att ttg ttc ggc tat tca ctc gct ttc gct 192Ile Val Ser Ile Ile Trp Ile Leu Phe Gly Tyr Ser Leu Ala Phe Ala 50 55 60ccc gga aac cca ttg atc ggg ggt ttg gat tgg atc gga ctg cac ggt 240Pro Gly Asn Pro Leu Ile Gly Gly Leu Asp Trp Ile Gly Leu His Gly65 70 75 80gtc ggt ttt gaa ccc aat gcc gat tat tcg gag acg atc ccg cat acg 288Val Gly Phe Glu Pro Asn Ala Asp Tyr Ser Glu Thr Ile Pro His Thr 85 90 95ctt tat atg atg ttc caa ttg aca ttt gcc gtc ctg acg act gcg att 336Leu Tyr Met Met Phe Gln Leu Thr Phe Ala Val Leu Thr Thr Ala Ile 100 105 110att tcc gga agc ttt gcg gag cgc atg cgc ttt ccg gcc ttt att ctt 384Ile Ser Gly Ser Phe Ala Glu Arg Met Arg Phe Pro Ala Phe Ile Leu 115 120 125ttc tcg gtg ctc tgg gcg gtt ttc gtt tat tcg cct gtc gcc cac tgg 432Phe Ser Val Leu Trp Ala Val Phe Val Tyr Ser Pro Val Ala His Trp 130 135 140gta tgg ggc gga ggc tgg ctt gac gag ctg gga gct att gat ttt gcc 480Val Trp Gly Gly Gly Trp Leu Asp Glu Leu Gly Ala Ile Asp Phe Ala145 150 155 160ggc gga aac gtc gtt cac att tct tcc ggg gta gcc ggt ctg gtc gtt 528Gly Gly Asn Val Val His Ile Ser Ser Gly Val Ala Gly Leu Val Val 165 170 175gcg att gtt ctc ggc aag cgg aaa aat atg agc gac acc gcc cct cac 576Ala Ile Val Leu Gly Lys Arg Lys Asn Met Ser Asp Thr Ala Pro His 180 185 190aac ctc ctt ctg acc ctg ctc ggg gcg act ttg atc tgg ttc gga tgg 624Asn Leu Leu Leu Thr Leu Leu Gly Ala Thr Leu Ile Trp Phe Gly Trp 195 200 205ttc ggt ttc aat gtc gga agc gcg ctg acg atc gat gaa gtc gcc atg 672Phe Gly Phe Asn Val Gly Ser Ala Leu Thr Ile Asp Glu Val Ala Met 210 215 220acc gct ttt atc aac acg aat acg gct gcg gcc gcg ggg ctg atc ggc 720Thr Ala Phe Ile Asn Thr Asn Thr Ala Ala Ala Ala Gly Leu Ile Gly225 230 235 240tgg att ttg gca gaa tgg ctg att aat aag aag ccg acg atg ctc ggc 768Trp Ile Leu Ala Glu Trp Leu Ile Asn Lys Lys Pro Thr Met Leu Gly 245 250 255gcc gtc tcg gga gcg att gcg gga ctc gtc gcg att acg cct ggc gca 816Ala Val Ser Gly Ala Ile Ala Gly Leu Val Ala Ile Thr Pro Gly Ala 260 265 270ggt ttt gtc acg cca ttc tca tct gta tgg atc ggg ctt atc gga gga 864Gly Phe Val Thr Pro Phe Ser Ser Val Trp Ile Gly Leu Ile Gly Gly 275 280 285atc gtc tgt ttc tgg ggg gtg ttt tcg ctc aag aaa aaa ttc ggg tac 912Ile Val Cys Phe Trp Gly Val Phe Ser Leu Lys Lys Lys Phe Gly Tyr 290 295 300gat gat gcg ctc gat gcc ttc gga ctt cac ggg ctc ggc ggg aca tgg 960Asp Asp Ala Leu Asp Ala Phe Gly Leu His Gly Leu Gly Gly Thr Trp305 310 315 320ggc gga atc gcg aca ggt ttg ttc gcg acg aca tct gtc aac gca gac 1008Gly Gly Ile Ala Thr Gly Leu Phe Ala Thr Thr Ser Val Asn Ala Asp 325 330 335ggt gcc aac ggg ctt ttt tac gga gac atc agt ctc ctt tgg aaa cag 1056Gly Ala Asn Gly Leu Phe Tyr Gly Asp Ile Ser Leu Leu Trp Lys Gln 340 345 350ctg att gcg att gct gca aca tat ttg ttt gtc gca atc gtc aca tac 1104Leu Ile Ala Ile Ala Ala Thr Tyr Leu Phe Val Ala Ile Val Thr Tyr 355 360 365gct att att aag ata gtc agt atc ttt ttt aaa ctc cgt gcc tca gag 1152Ala Ile Ile Lys Ile Val Ser Ile Phe Phe Lys Leu Arg Ala Ser Glu 370 375 380gat gaa gaa tca ttg ggc ctc gat ctt acg ctg cac ggc gaa aaa gcg 1200Asp Glu Glu Ser Leu Gly Leu Asp Leu Thr Leu His Gly Glu Lys Ala385 390 395 400tat caa gat taa 1212Tyr Gln Asp6403PRTBacillus licheniformis DSM 13 6Met Gln Met Gly Asp Thr Val Phe Met Phe Phe Cys Ala Leu Leu Val1 5 10 15Trp Leu Met Thr Pro Gly Ile Ala Leu Phe Tyr Gly Gly Leu Val Arg 20 25 30Ser Lys Asn Val Leu Ser Thr Val Met His Ser Leu Ser Ser Ile Ala 35 40 45Ile Val Ser Ile Ile Trp Ile Leu

Phe Gly Tyr Ser Leu Ala Phe Ala 50 55 60Pro Gly Asn Pro Leu Ile Gly Gly Leu Asp Trp Ile Gly Leu His Gly65 70 75 80Val Gly Phe Glu Pro Asn Ala Asp Tyr Ser Glu Thr Ile Pro His Thr 85 90 95Leu Tyr Met Met Phe Gln Leu Thr Phe Ala Val Leu Thr Thr Ala Ile 100 105 110Ile Ser Gly Ser Phe Ala Glu Arg Met Arg Phe Pro Ala Phe Ile Leu 115 120 125Phe Ser Val Leu Trp Ala Val Phe Val Tyr Ser Pro Val Ala His Trp 130 135 140Val Trp Gly Gly Gly Trp Leu Asp Glu Leu Gly Ala Ile Asp Phe Ala145 150 155 160Gly Gly Asn Val Val His Ile Ser Ser Gly Val Ala Gly Leu Val Val 165 170 175Ala Ile Val Leu Gly Lys Arg Lys Asn Met Ser Asp Thr Ala Pro His 180 185 190Asn Leu Leu Leu Thr Leu Leu Gly Ala Thr Leu Ile Trp Phe Gly Trp 195 200 205Phe Gly Phe Asn Val Gly Ser Ala Leu Thr Ile Asp Glu Val Ala Met 210 215 220Thr Ala Phe Ile Asn Thr Asn Thr Ala Ala Ala Ala Gly Leu Ile Gly225 230 235 240Trp Ile Leu Ala Glu Trp Leu Ile Asn Lys Lys Pro Thr Met Leu Gly 245 250 255Ala Val Ser Gly Ala Ile Ala Gly Leu Val Ala Ile Thr Pro Gly Ala 260 265 270Gly Phe Val Thr Pro Phe Ser Ser Val Trp Ile Gly Leu Ile Gly Gly 275 280 285Ile Val Cys Phe Trp Gly Val Phe Ser Leu Lys Lys Lys Phe Gly Tyr 290 295 300Asp Asp Ala Leu Asp Ala Phe Gly Leu His Gly Leu Gly Gly Thr Trp305 310 315 320Gly Gly Ile Ala Thr Gly Leu Phe Ala Thr Thr Ser Val Asn Ala Asp 325 330 335Gly Ala Asn Gly Leu Phe Tyr Gly Asp Ile Ser Leu Leu Trp Lys Gln 340 345 350Leu Ile Ala Ile Ala Ala Thr Tyr Leu Phe Val Ala Ile Val Thr Tyr 355 360 365Ala Ile Ile Lys Ile Val Ser Ile Phe Phe Lys Leu Arg Ala Ser Glu 370 375 380Asp Glu Glu Ser Leu Gly Leu Asp Leu Thr Leu His Gly Glu Lys Ala385 390 395 400Tyr Gln Asp71302DNABacillus licheniformis DSM 13gene(1)..(1302)htrA - serine protease Do (heat-shock protein) - BLi01390 7atg atg gaa tat aat aga gaa gat cag ctt gaa aca aaa aac gaa ctc 48Met Met Glu Tyr Asn Arg Glu Asp Gln Leu Glu Thr Lys Asn Glu Leu1 5 10 15atc gtc tcc gat caa aag ccc gac cag gcg ata aaa aaa gca gaa cag 96Ile Val Ser Asp Gln Lys Pro Asp Gln Ala Ile Lys Lys Ala Glu Gln 20 25 30caa aag cgg aga ctc tcg tgg atg acg ccg att ctc agc ggc atc atc 144Gln Lys Arg Arg Leu Ser Trp Met Thr Pro Ile Leu Ser Gly Ile Ile 35 40 45gga gga agc ctc gtt ctc ggc gtc acg aca tac cag cat agc aaa gac 192Gly Gly Ser Leu Val Leu Gly Val Thr Thr Tyr Gln His Ser Lys Asp 50 55 60gaa acg gcc aat aca tcc gtc cag acg caa aca aac gag acg tcg aac 240Glu Thr Ala Asn Thr Ser Val Gln Thr Gln Thr Asn Glu Thr Ser Asn65 70 75 80tca tcg gct tct tcc act aaa gcg aat acc gaa aaa ttg tcc agc acc 288Ser Ser Ala Ser Ser Thr Lys Ala Asn Thr Glu Lys Leu Ser Ser Thr 85 90 95aat tcg tcc gac att tct aca atg gtt gaa aat gta tct ccc gcc atc 336Asn Ser Ser Asp Ile Ser Thr Met Val Glu Asn Val Ser Pro Ala Ile 100 105 110gtc ggt att tca aac tat caa aaa caa aca gag agc ggt atg ttc ggt 384Val Gly Ile Ser Asn Tyr Gln Lys Gln Thr Glu Ser Gly Met Phe Gly 115 120 125ttt gaa agc aac agc agc agc tca agc gaa caa gaa aca ggc aca ggc 432Phe Glu Ser Asn Ser Ser Ser Ser Ser Glu Gln Glu Thr Gly Thr Gly 130 135 140tca ggt gtc atc tat aaa aaa gca aac ggc aaa gca tac att att aca 480Ser Gly Val Ile Tyr Lys Lys Ala Asn Gly Lys Ala Tyr Ile Ile Thr145 150 155 160aat aac cac gtt gtc gaa ggc gcg tct aaa ctg acg gtt tcc ctg tcc 528Asn Asn His Val Val Glu Gly Ala Ser Lys Leu Thr Val Ser Leu Ser 165 170 175aac ggc aaa gaa gtg gaa ggt aaa ttg ctg ggc agc gat tcg ttg aca 576Asn Gly Lys Glu Val Glu Gly Lys Leu Leu Gly Ser Asp Ser Leu Thr 180 185 190gac ctt gcc gtt gtt gaa att tcc gct gat cac gtg gaa aaa gtc gcc 624Asp Leu Ala Val Val Glu Ile Ser Ala Asp His Val Glu Lys Val Ala 195 200 205tct ttc ggc gat tcg tca agc ttg aaa gca ggc gaa tcg gtg atc gct 672Ser Phe Gly Asp Ser Ser Ser Leu Lys Ala Gly Glu Ser Val Ile Ala 210 215 220att gga aac ccg ctc ggc aaa gat ctg tcc cgc acg gtg aca caa ggc 720Ile Gly Asn Pro Leu Gly Lys Asp Leu Ser Arg Thr Val Thr Gln Gly225 230 235 240atc atc agc ggt gtt gat cgg acc gtt tcc gta gat aca tcc gca ggc 768Ile Ile Ser Gly Val Asp Arg Thr Val Ser Val Asp Thr Ser Ala Gly 245 250 255caa acc gag atg aac gtc att caa acc gac gcg gcc atc aac ccc ggc 816Gln Thr Glu Met Asn Val Ile Gln Thr Asp Ala Ala Ile Asn Pro Gly 260 265 270aac agc gga ggc gcc ctt ttg aat aca aag ggt gaa gtg atc ggc atc 864Asn Ser Gly Gly Ala Leu Leu Asn Thr Lys Gly Glu Val Ile Gly Ile 275 280 285aac agc atg aaa atc agt gaa agc gga gta gaa ggc atc gga ttt gcg 912Asn Ser Met Lys Ile Ser Glu Ser Gly Val Glu Gly Ile Gly Phe Ala 290 295 300att cca agc aat gat gtc aaa ccg att gca gaa gag ctg atg tct aaa 960Ile Pro Ser Asn Asp Val Lys Pro Ile Ala Glu Glu Leu Met Ser Lys305 310 315 320gga aaa att gag cgc ccg ttc atc gga atc ggc atg atg gat ctt gag 1008Gly Lys Ile Glu Arg Pro Phe Ile Gly Ile Gly Met Met Asp Leu Glu 325 330 335cag gtg ccg gaa aac tat caa aca ggc acc ctc gga tta agc ggc agc 1056Gln Val Pro Glu Asn Tyr Gln Thr Gly Thr Leu Gly Leu Ser Gly Ser 340 345 350cag tta aac aaa ggc gtc tac atc cgc cag gtc gct tca ggc tcc cca 1104Gln Leu Asn Lys Gly Val Tyr Ile Arg Gln Val Ala Ser Gly Ser Pro 355 360 365gct gaa aaa gcg ggc ctt aaa gag aac gat gtc atc gtc agc ttt aac 1152Ala Glu Lys Ala Gly Leu Lys Glu Asn Asp Val Ile Val Ser Phe Asn 370 375 380gga aaa gaa acc gat aca gga agc gcg ctg cgc aac ctt ctt tac aac 1200Gly Lys Glu Thr Asp Thr Gly Ser Ala Leu Arg Asn Leu Leu Tyr Asn385 390 395 400gat gca aaa atc ggg gat acg gta aag gtt acc ctc att cga aac ggc 1248Asp Ala Lys Ile Gly Asp Thr Val Lys Val Thr Leu Ile Arg Asn Gly 405 410 415aaa acc atg aca aaa caa atc acc ctt gac cag aaa gaa tcc gca tca 1296Lys Thr Met Thr Lys Gln Ile Thr Leu Asp Gln Lys Glu Ser Ala Ser 420 425 430tct taa 1302Ser 8433PRTBacillus licheniformis DSM 13 8Met Met Glu Tyr Asn Arg Glu Asp Gln Leu Glu Thr Lys Asn Glu Leu1 5 10 15Ile Val Ser Asp Gln Lys Pro Asp Gln Ala Ile Lys Lys Ala Glu Gln 20 25 30Gln Lys Arg Arg Leu Ser Trp Met Thr Pro Ile Leu Ser Gly Ile Ile 35 40 45Gly Gly Ser Leu Val Leu Gly Val Thr Thr Tyr Gln His Ser Lys Asp 50 55 60Glu Thr Ala Asn Thr Ser Val Gln Thr Gln Thr Asn Glu Thr Ser Asn65 70 75 80Ser Ser Ala Ser Ser Thr Lys Ala Asn Thr Glu Lys Leu Ser Ser Thr 85 90 95Asn Ser Ser Asp Ile Ser Thr Met Val Glu Asn Val Ser Pro Ala Ile 100 105 110Val Gly Ile Ser Asn Tyr Gln Lys Gln Thr Glu Ser Gly Met Phe Gly 115 120 125Phe Glu Ser Asn Ser Ser Ser Ser Ser Glu Gln Glu Thr Gly Thr Gly 130 135 140Ser Gly Val Ile Tyr Lys Lys Ala Asn Gly Lys Ala Tyr Ile Ile Thr145 150 155 160Asn Asn His Val Val Glu Gly Ala Ser Lys Leu Thr Val Ser Leu Ser 165 170 175Asn Gly Lys Glu Val Glu Gly Lys Leu Leu Gly Ser Asp Ser Leu Thr 180 185 190Asp Leu Ala Val Val Glu Ile Ser Ala Asp His Val Glu Lys Val Ala 195 200 205Ser Phe Gly Asp Ser Ser Ser Leu Lys Ala Gly Glu Ser Val Ile Ala 210 215 220Ile Gly Asn Pro Leu Gly Lys Asp Leu Ser Arg Thr Val Thr Gln Gly225 230 235 240Ile Ile Ser Gly Val Asp Arg Thr Val Ser Val Asp Thr Ser Ala Gly 245 250 255Gln Thr Glu Met Asn Val Ile Gln Thr Asp Ala Ala Ile Asn Pro Gly 260 265 270Asn Ser Gly Gly Ala Leu Leu Asn Thr Lys Gly Glu Val Ile Gly Ile 275 280 285Asn Ser Met Lys Ile Ser Glu Ser Gly Val Glu Gly Ile Gly Phe Ala 290 295 300Ile Pro Ser Asn Asp Val Lys Pro Ile Ala Glu Glu Leu Met Ser Lys305 310 315 320Gly Lys Ile Glu Arg Pro Phe Ile Gly Ile Gly Met Met Asp Leu Glu 325 330 335Gln Val Pro Glu Asn Tyr Gln Thr Gly Thr Leu Gly Leu Ser Gly Ser 340 345 350Gln Leu Asn Lys Gly Val Tyr Ile Arg Gln Val Ala Ser Gly Ser Pro 355 360 365Ala Glu Lys Ala Gly Leu Lys Glu Asn Asp Val Ile Val Ser Phe Asn 370 375 380Gly Lys Glu Thr Asp Thr Gly Ser Ala Leu Arg Asn Leu Leu Tyr Asn385 390 395 400Asp Ala Lys Ile Gly Asp Thr Val Lys Val Thr Leu Ile Arg Asn Gly 405 410 415Lys Thr Met Thr Lys Gln Ile Thr Leu Asp Gln Lys Glu Ser Ala Ser 420 425 430Ser91215DNABacillus licheniformis DSM 13gene(1)..(1215)putative protein - putative serine protease- BLi00301 9ttg aaa tcg aaa tgg agt gca atg gtg gtt att gcc ggt ctt tta ttg 48Leu Lys Ser Lys Trp Ser Ala Met Val Val Ile Ala Gly Leu Leu Leu1 5 10 15ctg gcc gga tgc ggt gca ctg aag gag gct gat ccc ccg cgc ggc aga 96Leu Ala Gly Cys Gly Ala Leu Lys Glu Ala Asp Pro Pro Arg Gly Arg 20 25 30tca gcg caa aaa acg gaa gcc tcg ttt tct gaa gct gaa cag cga ttt 144Ser Ala Gln Lys Thr Glu Ala Ser Phe Ser Glu Ala Glu Gln Arg Phe 35 40 45gcg ctc gcc ttg ttt caa gac atg ata aaa gaa gaa ggg agc cgg aaa 192Ala Leu Ala Leu Phe Gln Asp Met Ile Lys Glu Glu Gly Ser Arg Lys 50 55 60aac atc ttc ctc tcg cct tac agt att cag cag gca ctt ttg atg acg 240Asn Ile Phe Leu Ser Pro Tyr Ser Ile Gln Gln Ala Leu Leu Met Thr65 70 75 80gca aac ggt gcc gcg gga gac agc aga aag gaa ctg ctc agc act tta 288Ala Asn Gly Ala Ala Gly Asp Ser Arg Lys Glu Leu Leu Ser Thr Leu 85 90 95cat ctc agc cag gcg gat atg gca tcg atc aac ggg ata tca aaa tct 336His Leu Ser Gln Ala Asp Met Ala Ser Ile Asn Gly Ile Ser Lys Ser 100 105 110gtt aat cgt tct ctt gaa acg ctg cct cac ggt gaa ttt tca tca gct 384Val Asn Arg Ser Leu Glu Thr Leu Pro His Gly Glu Phe Ser Ser Ala 115 120 125aat tcg ata tgg acc aaa agg gag ctg aaa gaa agc ttt caa gac gcg 432Asn Ser Ile Trp Thr Lys Arg Glu Leu Lys Glu Ser Phe Gln Asp Ala 130 135 140atc aaa ggc ctt ggc gac gcc tat cag ctt caa att gat cct aat cgg 480Ile Lys Gly Leu Gly Asp Ala Tyr Gln Leu Gln Ile Asp Pro Asn Arg145 150 155 160gca aca gag gaa atc aac agc tgg gtc aaa aag aaa acg aaa gac cgc 528Ala Thr Glu Glu Ile Asn Ser Trp Val Lys Lys Lys Thr Lys Asp Arg 165 170 175atc gat cag atc gtt gat aaa gtc tcg tcg aat acg att gcc tac atc 576Ile Asp Gln Ile Val Asp Lys Val Ser Ser Asn Thr Ile Ala Tyr Ile 180 185 190atc aat gcc gtc tat ttt aaa gaa agc tgg aag cac cca ttt gat cct 624Ile Asn Ala Val Tyr Phe Lys Glu Ser Trp Lys His Pro Phe Asp Pro 195 200 205cac atg aca gcg gaa cag cca ttt tat tca gcg gac ggc tcc gca aaa 672His Met Thr Ala Glu Gln Pro Phe Tyr Ser Ala Asp Gly Ser Ala Lys 210 215 220aaa cac ccc atg atg acg cag aca aac cgc ttt cct tat ctg gaa aac 720Lys His Pro Met Met Thr Gln Thr Asn Arg Phe Pro Tyr Leu Glu Asn225 230 235 240gaa cat ttt caa gcc gtc aag ctg cca ttt caa gac gaa ggc ctg tca 768Glu His Phe Gln Ala Val Lys Leu Pro Phe Gln Asp Glu Gly Leu Ser 245 250 255cta gcc gtt ttt ctg ccg aaa aaa gcg gca aat cta gaa cag ctt ctg 816Leu Ala Val Phe Leu Pro Lys Lys Ala Ala Asn Leu Glu Gln Leu Leu 260 265 270act ggc atg acg cat gat caa tgg cgc gcc ttg caa aag ggc tgg acg 864Thr Gly Met Thr His Asp Gln Trp Arg Ala Leu Gln Lys Gly Trp Thr 275 280 285atg aaa cag gtg gag ctc aag ctg ccg cgc ttc tca ttt caa aca gac 912Met Lys Gln Val Glu Leu Lys Leu Pro Arg Phe Ser Phe Gln Thr Asp 290 295 300tac atg tta aat gaa ccg ctt aaa cgc ctt gga atg aaa aac gtg ttt 960Tyr Met Leu Asn Glu Pro Leu Lys Arg Leu Gly Met Lys Asn Val Phe305 310 315 320agc agc gcg aat ttt tca aat atg ttt acc ggg cat ggc gcc gct caa 1008Ser Ser Ala Asn Phe Ser Asn Met Phe Thr Gly His Gly Ala Ala Gln 325 330 335att aat aaa gtc aga cat aaa aca ttt atc aaa gtg gat gaa gcg ggc 1056Ile Asn Lys Val Arg His Lys Thr Phe Ile Lys Val Asp Glu Ala Gly 340 345 350act gaa gca tcc gcg gca acg gcg gtt gaa atc atc gaa tca gcc ccc 1104Thr Glu Ala Ser Ala Ala Thr Ala Val Glu Ile Ile Glu Ser Ala Pro 355 360 365gtc cct gaa gtg acc atg act gca gac cac ccg ttt tat ttt gcg atc 1152Val Pro Glu Val Thr Met Thr Ala Asp His Pro Phe Tyr Phe Ala Ile 370 375 380gtt gat gaa gca tcc gga atg att ctc ttt ttg ggc agt gtg gca gag 1200Val Asp Glu Ala Ser Gly Met Ile Leu Phe Leu Gly Ser Val Ala Glu385 390 395 400ccg aaa gat gat taa 1215Pro Lys Asp Asp10404PRTBacillus licheniformis DSM 13misc_feature(1)..(1)Xaa can be Leu or Met. 10Xaa Lys Ser Lys Trp Ser Ala Met Val Val Ile Ala Gly Leu Leu Leu1 5 10 15Leu Ala Gly Cys Gly Ala Leu Lys Glu Ala Asp Pro Pro Arg Gly Arg 20 25 30Ser Ala Gln Lys Thr Glu Ala Ser Phe Ser Glu Ala Glu Gln Arg Phe 35 40 45Ala Leu Ala Leu Phe Gln Asp Met Ile Lys Glu Glu Gly Ser Arg Lys 50 55 60Asn Ile Phe Leu Ser Pro Tyr Ser Ile Gln Gln Ala Leu Leu Met Thr65 70 75 80Ala Asn Gly Ala Ala Gly Asp Ser Arg Lys Glu Leu Leu Ser Thr Leu 85 90 95His Leu Ser Gln Ala Asp Met Ala Ser Ile Asn Gly Ile Ser Lys Ser 100 105 110Val Asn Arg Ser Leu Glu Thr Leu Pro His Gly Glu Phe Ser Ser Ala 115 120 125Asn Ser Ile Trp Thr Lys Arg Glu Leu Lys Glu Ser Phe Gln Asp Ala 130 135 140Ile Lys Gly Leu Gly Asp Ala Tyr Gln Leu Gln Ile Asp Pro Asn Arg145 150 155 160Ala Thr Glu Glu Ile Asn Ser Trp Val Lys Lys Lys Thr Lys Asp Arg 165 170 175Ile Asp Gln Ile Val Asp Lys Val Ser Ser Asn Thr Ile Ala Tyr Ile 180 185 190Ile Asn Ala Val Tyr Phe Lys Glu Ser Trp Lys His Pro Phe Asp Pro 195 200 205His Met Thr Ala Glu Gln Pro Phe Tyr Ser Ala Asp Gly Ser Ala Lys 210 215 220Lys His Pro Met Met Thr Gln Thr Asn Arg Phe Pro Tyr Leu Glu Asn225 230 235 240Glu His Phe Gln Ala Val Lys Leu Pro Phe Gln Asp Glu Gly Leu Ser 245 250 255Leu Ala Val Phe Leu Pro Lys Lys Ala Ala Asn Leu Glu Gln Leu Leu 260 265 270Thr Gly Met Thr His Asp Gln Trp Arg Ala Leu Gln Lys Gly Trp Thr 275

280 285Met Lys Gln Val Glu Leu Lys Leu Pro Arg Phe Ser Phe Gln Thr Asp 290 295 300Tyr Met Leu Asn Glu Pro Leu Lys Arg Leu Gly Met Lys Asn Val Phe305 310 315 320Ser Ser Ala Asn Phe Ser Asn Met Phe Thr Gly His Gly Ala Ala Gln 325 330 335Ile Asn Lys Val Arg His Lys Thr Phe Ile Lys Val Asp Glu Ala Gly 340 345 350Thr Glu Ala Ser Ala Ala Thr Ala Val Glu Ile Ile Glu Ser Ala Pro 355 360 365Val Pro Glu Val Thr Met Thr Ala Asp His Pro Phe Tyr Phe Ala Ile 370 375 380Val Asp Glu Ala Ser Gly Met Ile Leu Phe Leu Gly Ser Val Ala Glu385 390 395 400Pro Lys Asp Asp11408DNABacillus licheniformis DSM 13gene(1)..(408)glnR - transcriptional repressor of the glutamine synthetase gene- BLi01992 11atg agt gat aaa att cgc cgc tca atg ccc tta ttt ccg ata ggt atc 48Met Ser Asp Lys Ile Arg Arg Ser Met Pro Leu Phe Pro Ile Gly Ile1 5 10 15gtc atg cag ctg act gaa ctg tcc gca agg caa att cgg tat tat gag 96Val Met Gln Leu Thr Glu Leu Ser Ala Arg Gln Ile Arg Tyr Tyr Glu 20 25 30gaa aat gga ctg gta ttt cct gca aga agc gaa ggg aac aga cgt tta 144Glu Asn Gly Leu Val Phe Pro Ala Arg Ser Glu Gly Asn Arg Arg Leu 35 40 45ttt tca ttc cat gat gta gat aag ctg ctc gaa att aaa gat ttg atc 192Phe Ser Phe His Asp Val Asp Lys Leu Leu Glu Ile Lys Asp Leu Ile 50 55 60gaa caa ggt gta aac atg gcg ggg atc aaa caa att ttc gcc aaa aca 240Glu Gln Gly Val Asn Met Ala Gly Ile Lys Gln Ile Phe Ala Lys Thr65 70 75 80gaa aac gaa caa aaa gaa gac aaa caa gac aag ccg aaa aaa gca gag 288Glu Asn Glu Gln Lys Glu Asp Lys Gln Asp Lys Pro Lys Lys Ala Glu 85 90 95aag cac aac ttg tcg gat gaa gaa ctg aga aaa ctg ctg aaa aac gag 336Lys His Asn Leu Ser Asp Glu Glu Leu Arg Lys Leu Leu Lys Asn Glu 100 105 110ctc atc caa gca ggc cgt ttc caa aga ggg acg aca ttc cgc cag gga 384Leu Ile Gln Ala Gly Arg Phe Gln Arg Gly Thr Thr Phe Arg Gln Gly 115 120 125gac atg tcc agg ttt ttt cat taa 408Asp Met Ser Arg Phe Phe His 130 13512135PRTBacillus licheniformis DSM 13 12Met Ser Asp Lys Ile Arg Arg Ser Met Pro Leu Phe Pro Ile Gly Ile1 5 10 15Val Met Gln Leu Thr Glu Leu Ser Ala Arg Gln Ile Arg Tyr Tyr Glu 20 25 30Glu Asn Gly Leu Val Phe Pro Ala Arg Ser Glu Gly Asn Arg Arg Leu 35 40 45Phe Ser Phe His Asp Val Asp Lys Leu Leu Glu Ile Lys Asp Leu Ile 50 55 60Glu Gln Gly Val Asn Met Ala Gly Ile Lys Gln Ile Phe Ala Lys Thr65 70 75 80Glu Asn Glu Gln Lys Glu Asp Lys Gln Asp Lys Pro Lys Lys Ala Glu 85 90 95Lys His Asn Leu Ser Asp Glu Glu Leu Arg Lys Leu Leu Lys Asn Glu 100 105 110Leu Ile Gln Ala Gly Arg Phe Gln Arg Gly Thr Thr Phe Arg Gln Gly 115 120 125Asp Met Ser Arg Phe Phe His 130 135131335DNABacillus licheniformis DSM 13gene(1)..(1335)glnA - glutamine synthetase - BLi01993 13atg gca aag tat aca aga gac gac att gtt aaa cta gta aat gag gag 48Met Ala Lys Tyr Thr Arg Asp Asp Ile Val Lys Leu Val Asn Glu Glu1 5 10 15aat gta aaa tat att cgc ctt cag ttt aca gac att ctt gga acg att 96Asn Val Lys Tyr Ile Arg Leu Gln Phe Thr Asp Ile Leu Gly Thr Ile 20 25 30aaa aac gtt gaa att cct gtg agc cag ctg gaa aaa gca ctt gat aat 144Lys Asn Val Glu Ile Pro Val Ser Gln Leu Glu Lys Ala Leu Asp Asn 35 40 45aaa gtc atg ttc gac ggc tct tca atc gag gga ttc gtc cgc att gaa 192Lys Val Met Phe Asp Gly Ser Ser Ile Glu Gly Phe Val Arg Ile Glu 50 55 60gaa tcc gat atg tac tta tac cct gac tta gat aca ttt gtg att ttc 240Glu Ser Asp Met Tyr Leu Tyr Pro Asp Leu Asp Thr Phe Val Ile Phe65 70 75 80cca tgg acc gcc gaa aaa ggt aaa gta gcc cgc ttc atc tgc gat att 288Pro Trp Thr Ala Glu Lys Gly Lys Val Ala Arg Phe Ile Cys Asp Ile 85 90 95tat aat ccg gac ggc acg ccg ttt gaa ggt gat ccg cgc aac aac tta 336Tyr Asn Pro Asp Gly Thr Pro Phe Glu Gly Asp Pro Arg Asn Asn Leu 100 105 110aaa cgc att ttg aaa gaa atg gaa gac atg ggc ttc tca gat ttc aat 384Lys Arg Ile Leu Lys Glu Met Glu Asp Met Gly Phe Ser Asp Phe Asn 115 120 125ctt gga cca gag ccg gaa ttc ttc ctg ttc aag cta gac gaa aaa ggc 432Leu Gly Pro Glu Pro Glu Phe Phe Leu Phe Lys Leu Asp Glu Lys Gly 130 135 140gag cct acg ctt gaa ctg aac gac aaa ggc gga tat ttc gac ctt gcg 480Glu Pro Thr Leu Glu Leu Asn Asp Lys Gly Gly Tyr Phe Asp Leu Ala145 150 155 160ccg act gat ctt ggc gaa aac tgc cgc cgc gac atc gta ctt gag ctc 528Pro Thr Asp Leu Gly Glu Asn Cys Arg Arg Asp Ile Val Leu Glu Leu 165 170 175gaa gaa atg ggc ttt gaa atc gaa gcg tct cac cac gaa gtt gca cct 576Glu Glu Met Gly Phe Glu Ile Glu Ala Ser His His Glu Val Ala Pro 180 185 190gga cag cac gaa atc gac ttc aaa tac gcc gga gcc atc cgc gct tgc 624Gly Gln His Glu Ile Asp Phe Lys Tyr Ala Gly Ala Ile Arg Ala Cys 195 200 205gat gac atc caa acg ttc aag ctc gtt gtt aaa aca atc gcg cgc aaa 672Asp Asp Ile Gln Thr Phe Lys Leu Val Val Lys Thr Ile Ala Arg Lys 210 215 220cac ggc ctg cac gca aca ttc atg cca aaa ccg ttg ttc ggc gta aac 720His Gly Leu His Ala Thr Phe Met Pro Lys Pro Leu Phe Gly Val Asn225 230 235 240gga tca ggc atg cac tgc aat cta tca ctc ttc cga aac ggc gcc aac 768Gly Ser Gly Met His Cys Asn Leu Ser Leu Phe Arg Asn Gly Ala Asn 245 250 255gcc ttc ttt gac aaa gac gcc gat ctt cag ctg agt gaa acg gca aag 816Ala Phe Phe Asp Lys Asp Ala Asp Leu Gln Leu Ser Glu Thr Ala Lys 260 265 270caa ttc atc gcc ggc atc gtc aag cac gca aca agc ttt aca gct gtg 864Gln Phe Ile Ala Gly Ile Val Lys His Ala Thr Ser Phe Thr Ala Val 275 280 285aca aac ccg act gtc aac tct tac aag cgc ctt gtt cca ggc tat gaa 912Thr Asn Pro Thr Val Asn Ser Tyr Lys Arg Leu Val Pro Gly Tyr Glu 290 295 300gca cct tgc tat gtg gca tgg agc gcg caa aac aga agc ccg tta atc 960Ala Pro Cys Tyr Val Ala Trp Ser Ala Gln Asn Arg Ser Pro Leu Ile305 310 315 320cgc atc ccg gct tcc cgc ggc atc agc acg cgc gta gaa gtc aga agc 1008Arg Ile Pro Ala Ser Arg Gly Ile Ser Thr Arg Val Glu Val Arg Ser 325 330 335gta gac cct tct gcg aac ccg tac ctt gcc ttg agc gta ctt ctt gca 1056Val Asp Pro Ser Ala Asn Pro Tyr Leu Ala Leu Ser Val Leu Leu Ala 340 345 350gca gga ctt gac ggc atc aaa aac aag ctg gaa gca ccg gct ccg atc 1104Ala Gly Leu Asp Gly Ile Lys Asn Lys Leu Glu Ala Pro Ala Pro Ile 355 360 365gac cgc aac atc tat gtc atg aca aaa gaa gag cgc ctc gaa aac gga 1152Asp Arg Asn Ile Tyr Val Met Thr Lys Glu Glu Arg Leu Glu Asn Gly 370 375 380atc gtc gac ctt cct gcg aca ctc gca gac gct ctc gaa gaa ttc aaa 1200Ile Val Asp Leu Pro Ala Thr Leu Ala Asp Ala Leu Glu Glu Phe Lys385 390 395 400tca aac gaa gtc atg gtc aaa gcc ctc ggc gac cat cta ttc gaa cac 1248Ser Asn Glu Val Met Val Lys Ala Leu Gly Asp His Leu Phe Glu His 405 410 415ttc gtc gaa gca aaa gaa atc gaa tgg gat atg ttc cgc aca caa gtc 1296Phe Val Glu Ala Lys Glu Ile Glu Trp Asp Met Phe Arg Thr Gln Val 420 425 430cat cct tgg gag cgc gag cag tat atg tct cag tat taa 1335His Pro Trp Glu Arg Glu Gln Tyr Met Ser Gln Tyr 435 44014444PRTBacillus licheniformis DSM 13 14Met Ala Lys Tyr Thr Arg Asp Asp Ile Val Lys Leu Val Asn Glu Glu1 5 10 15Asn Val Lys Tyr Ile Arg Leu Gln Phe Thr Asp Ile Leu Gly Thr Ile 20 25 30Lys Asn Val Glu Ile Pro Val Ser Gln Leu Glu Lys Ala Leu Asp Asn 35 40 45Lys Val Met Phe Asp Gly Ser Ser Ile Glu Gly Phe Val Arg Ile Glu 50 55 60Glu Ser Asp Met Tyr Leu Tyr Pro Asp Leu Asp Thr Phe Val Ile Phe65 70 75 80Pro Trp Thr Ala Glu Lys Gly Lys Val Ala Arg Phe Ile Cys Asp Ile 85 90 95Tyr Asn Pro Asp Gly Thr Pro Phe Glu Gly Asp Pro Arg Asn Asn Leu 100 105 110Lys Arg Ile Leu Lys Glu Met Glu Asp Met Gly Phe Ser Asp Phe Asn 115 120 125Leu Gly Pro Glu Pro Glu Phe Phe Leu Phe Lys Leu Asp Glu Lys Gly 130 135 140Glu Pro Thr Leu Glu Leu Asn Asp Lys Gly Gly Tyr Phe Asp Leu Ala145 150 155 160Pro Thr Asp Leu Gly Glu Asn Cys Arg Arg Asp Ile Val Leu Glu Leu 165 170 175Glu Glu Met Gly Phe Glu Ile Glu Ala Ser His His Glu Val Ala Pro 180 185 190Gly Gln His Glu Ile Asp Phe Lys Tyr Ala Gly Ala Ile Arg Ala Cys 195 200 205Asp Asp Ile Gln Thr Phe Lys Leu Val Val Lys Thr Ile Ala Arg Lys 210 215 220His Gly Leu His Ala Thr Phe Met Pro Lys Pro Leu Phe Gly Val Asn225 230 235 240Gly Ser Gly Met His Cys Asn Leu Ser Leu Phe Arg Asn Gly Ala Asn 245 250 255Ala Phe Phe Asp Lys Asp Ala Asp Leu Gln Leu Ser Glu Thr Ala Lys 260 265 270Gln Phe Ile Ala Gly Ile Val Lys His Ala Thr Ser Phe Thr Ala Val 275 280 285Thr Asn Pro Thr Val Asn Ser Tyr Lys Arg Leu Val Pro Gly Tyr Glu 290 295 300Ala Pro Cys Tyr Val Ala Trp Ser Ala Gln Asn Arg Ser Pro Leu Ile305 310 315 320Arg Ile Pro Ala Ser Arg Gly Ile Ser Thr Arg Val Glu Val Arg Ser 325 330 335Val Asp Pro Ser Ala Asn Pro Tyr Leu Ala Leu Ser Val Leu Leu Ala 340 345 350Ala Gly Leu Asp Gly Ile Lys Asn Lys Leu Glu Ala Pro Ala Pro Ile 355 360 365Asp Arg Asn Ile Tyr Val Met Thr Lys Glu Glu Arg Leu Glu Asn Gly 370 375 380Ile Val Asp Leu Pro Ala Thr Leu Ala Asp Ala Leu Glu Glu Phe Lys385 390 395 400Ser Asn Glu Val Met Val Lys Ala Leu Gly Asp His Leu Phe Glu His 405 410 415Phe Val Glu Ala Lys Glu Ile Glu Trp Asp Met Phe Arg Thr Gln Val 420 425 430His Pro Trp Glu Arg Glu Gln Tyr Met Ser Gln Tyr 435 440151476DNABacillus licheniformis DSM 13gene(1)..(1476)hypothetical protein - close homolog to DhaS aldehyde dehydrogenase - BLi03994 15atg aat gtt caa act cca tcg gaa aaa ctc gta aaa cct ttt gac tgt 48Met Asn Val Gln Thr Pro Ser Glu Lys Leu Val Lys Pro Phe Asp Cys1 5 10 15cag cac tac ata aac ggc agg tac ttt cct tca gag aaa ggc agc acg 96Gln His Tyr Ile Asn Gly Arg Tyr Phe Pro Ser Glu Lys Gly Ser Thr 20 25 30ttt gac aac atc aat ccg gct aca cag gaa att atc gga agc gtt gcg 144Phe Asp Asn Ile Asn Pro Ala Thr Gln Glu Ile Ile Gly Ser Val Ala 35 40 45gaa gga ggc aaa aaa gag gtc gat ctt gca gtc gcg gca gcc agg cgt 192Glu Gly Gly Lys Lys Glu Val Asp Leu Ala Val Ala Ala Ala Arg Arg 50 55 60gca tta aac gga cgc tgg aaa aac atg acc gct gac gaa agg att cgg 240Ala Leu Asn Gly Arg Trp Lys Asn Met Thr Ala Asp Glu Arg Ile Arg65 70 75 80atc atc aga aag gtc ggc gac atc ata ctt gag cgt aaa gac gaa ctg 288Ile Ile Arg Lys Val Gly Asp Ile Ile Leu Glu Arg Lys Asp Glu Leu 85 90 95gcg agg ctt gag aca ttg gat acg gga aaa ccg ctt tcg ctc tca agc 336Ala Arg Leu Glu Thr Leu Asp Thr Gly Lys Pro Leu Ser Leu Ser Ser 100 105 110act ttg gat att cca cgt gcc gct ttc aac ttt cat ttc ttt gcg gat 384Thr Leu Asp Ile Pro Arg Ala Ala Phe Asn Phe His Phe Phe Ala Asp 115 120 125ttt atg aga gga gcc gga acg gaa gct tat caa act gac gat ctt gca 432Phe Met Arg Gly Ala Gly Thr Glu Ala Tyr Gln Thr Asp Asp Leu Ala 130 135 140atc aat tat gcg att cgc cgt ccg gtc ggc gtc gtc ggg ctg atc aat 480Ile Asn Tyr Ala Ile Arg Arg Pro Val Gly Val Val Gly Leu Ile Asn145 150 155 160ccg tgg aac ttg ccg ctt ctt ttg ctg acg tgg aaa ttg gcg cct tgc 528Pro Trp Asn Leu Pro Leu Leu Leu Leu Thr Trp Lys Leu Ala Pro Cys 165 170 175ctg gcg gcg gga aat aca gcc gtt atc aaa ccg gct gaa ctg acg ccg 576Leu Ala Ala Gly Asn Thr Ala Val Ile Lys Pro Ala Glu Leu Thr Pro 180 185 190atg aca gcc acc ctt ctc ggg gaa atc tgc cat gat gca ggg atg cct 624Met Thr Ala Thr Leu Leu Gly Glu Ile Cys His Asp Ala Gly Met Pro 195 200 205gac gga gtc gtt aat atc gtc cac gga ttt gga cca gat tcc gcc ggt 672Asp Gly Val Val Asn Ile Val His Gly Phe Gly Pro Asp Ser Ala Gly 210 215 220gct gcg ctt tct gag cat ccc gat gtc gat gcg ata tct ttc acc gga 720Ala Ala Leu Ser Glu His Pro Asp Val Asp Ala Ile Ser Phe Thr Gly225 230 235 240gaa acg aca acc ggc aaa gtc att atg gaa gct gct tcg aaa aca tta 768Glu Thr Thr Thr Gly Lys Val Ile Met Glu Ala Ala Ser Lys Thr Leu 245 250 255aag aaa ctt tcc ttt gaa ttg gga ggg aag aac ccg aac att att ttt 816Lys Lys Leu Ser Phe Glu Leu Gly Gly Lys Asn Pro Asn Ile Ile Phe 260 265 270gcc gat gcc gac atg gat gaa gcc gtc agc acc tca ttg aaa tcg agt 864Ala Asp Ala Asp Met Asp Glu Ala Val Ser Thr Ser Leu Lys Ser Ser 275 280 285ttc gtc aac cag ggg gaa gta tgt atg agc ggc tcc cgc atc tat gtc 912Phe Val Asn Gln Gly Glu Val Cys Met Ser Gly Ser Arg Ile Tyr Val 290 295 300gag agg gaa gcg tat gac gaa ttt ctg aac aag ttc gtt gag aaa acg 960Glu Arg Glu Ala Tyr Asp Glu Phe Leu Asn Lys Phe Val Glu Lys Thr305 310 315 320aaa aag ctc aaa gtc ggc gat cct ttt gat ccg gac acg aat atc ggc 1008Lys Lys Leu Lys Val Gly Asp Pro Phe Asp Pro Asp Thr Asn Ile Gly 325 330 335gcc ttg att tct tct gaa cac aca gag cgt gtg atg aac tac atc gaa 1056Ala Leu Ile Ser Ser Glu His Thr Glu Arg Val Met Asn Tyr Ile Glu 340 345 350ttg gcc aaa cgg gaa ggc ggc aat att tta acc gga gga aaa cgc cct 1104Leu Ala Lys Arg Glu Gly Gly Asn Ile Leu Thr Gly Gly Lys Arg Pro 355 360 365gaa gga cag gaa tcc ggc tgc ttt ctt gaa ccg acg att atc acc gga 1152Glu Gly Gln Glu Ser Gly Cys Phe Leu Glu Pro Thr Ile Ile Thr Gly 370 375 380ctt tca cgc aat tcc agg ctc atc aaa gaa gaa atc ttc ggt ccg gtc 1200Leu Ser Arg Asn Ser Arg Leu Ile Lys Glu Glu Ile Phe Gly Pro Val385 390 395 400gta acg gtg att ccg ttc gac gat gag gag gaa gtg att gca cag gct 1248Val Thr Val Ile Pro Phe Asp Asp Glu Glu Glu Val Ile Ala Gln Ala 405 410 415aac gac aca cat tac ggg tta agc gcg acc tta tgg acg aac gac ctc 1296Asn Asp Thr His Tyr Gly Leu Ser Ala Thr Leu Trp Thr Asn Asp Leu 420 425 430cgc cgg gcg cac cgc gta gcc ggg caa atc gaa tcg ggc atg gtg tgg 1344Arg Arg Ala His Arg Val Ala Gly Gln Ile Glu Ser Gly Met Val Trp 435 440 445gtc aac agc tgg ttt ctc cgc gat ctt cgg acg ccg ttt ggc ggc atg 1392Val Asn Ser Trp Phe Leu Arg Asp Leu Arg Thr Pro Phe Gly Gly Met 450 455 460aaa cag agc gga ctc gga cga gaa gga ggc att cac agc ctc gaa ttc 1440Lys Gln Ser Gly Leu Gly Arg Glu Gly Gly Ile His Ser Leu Glu Phe465 470 475 480ttc agc gaa tta

acc aat att tgc atc aag ctg taa 1476Phe Ser Glu Leu Thr Asn Ile Cys Ile Lys Leu 485 49016491PRTBacillus licheniformis DSM 13 16Met Asn Val Gln Thr Pro Ser Glu Lys Leu Val Lys Pro Phe Asp Cys1 5 10 15Gln His Tyr Ile Asn Gly Arg Tyr Phe Pro Ser Glu Lys Gly Ser Thr 20 25 30Phe Asp Asn Ile Asn Pro Ala Thr Gln Glu Ile Ile Gly Ser Val Ala 35 40 45Glu Gly Gly Lys Lys Glu Val Asp Leu Ala Val Ala Ala Ala Arg Arg 50 55 60Ala Leu Asn Gly Arg Trp Lys Asn Met Thr Ala Asp Glu Arg Ile Arg65 70 75 80Ile Ile Arg Lys Val Gly Asp Ile Ile Leu Glu Arg Lys Asp Glu Leu 85 90 95Ala Arg Leu Glu Thr Leu Asp Thr Gly Lys Pro Leu Ser Leu Ser Ser 100 105 110Thr Leu Asp Ile Pro Arg Ala Ala Phe Asn Phe His Phe Phe Ala Asp 115 120 125Phe Met Arg Gly Ala Gly Thr Glu Ala Tyr Gln Thr Asp Asp Leu Ala 130 135 140Ile Asn Tyr Ala Ile Arg Arg Pro Val Gly Val Val Gly Leu Ile Asn145 150 155 160Pro Trp Asn Leu Pro Leu Leu Leu Leu Thr Trp Lys Leu Ala Pro Cys 165 170 175Leu Ala Ala Gly Asn Thr Ala Val Ile Lys Pro Ala Glu Leu Thr Pro 180 185 190Met Thr Ala Thr Leu Leu Gly Glu Ile Cys His Asp Ala Gly Met Pro 195 200 205Asp Gly Val Val Asn Ile Val His Gly Phe Gly Pro Asp Ser Ala Gly 210 215 220Ala Ala Leu Ser Glu His Pro Asp Val Asp Ala Ile Ser Phe Thr Gly225 230 235 240Glu Thr Thr Thr Gly Lys Val Ile Met Glu Ala Ala Ser Lys Thr Leu 245 250 255Lys Lys Leu Ser Phe Glu Leu Gly Gly Lys Asn Pro Asn Ile Ile Phe 260 265 270Ala Asp Ala Asp Met Asp Glu Ala Val Ser Thr Ser Leu Lys Ser Ser 275 280 285Phe Val Asn Gln Gly Glu Val Cys Met Ser Gly Ser Arg Ile Tyr Val 290 295 300Glu Arg Glu Ala Tyr Asp Glu Phe Leu Asn Lys Phe Val Glu Lys Thr305 310 315 320Lys Lys Leu Lys Val Gly Asp Pro Phe Asp Pro Asp Thr Asn Ile Gly 325 330 335Ala Leu Ile Ser Ser Glu His Thr Glu Arg Val Met Asn Tyr Ile Glu 340 345 350Leu Ala Lys Arg Glu Gly Gly Asn Ile Leu Thr Gly Gly Lys Arg Pro 355 360 365Glu Gly Gln Glu Ser Gly Cys Phe Leu Glu Pro Thr Ile Ile Thr Gly 370 375 380Leu Ser Arg Asn Ser Arg Leu Ile Lys Glu Glu Ile Phe Gly Pro Val385 390 395 400Val Thr Val Ile Pro Phe Asp Asp Glu Glu Glu Val Ile Ala Gln Ala 405 410 415Asn Asp Thr His Tyr Gly Leu Ser Ala Thr Leu Trp Thr Asn Asp Leu 420 425 430Arg Arg Ala His Arg Val Ala Gly Gln Ile Glu Ser Gly Met Val Trp 435 440 445Val Asn Ser Trp Phe Leu Arg Asp Leu Arg Thr Pro Phe Gly Gly Met 450 455 460Lys Gln Ser Gly Leu Gly Arg Glu Gly Gly Ile His Ser Leu Glu Phe465 470 475 480Phe Ser Glu Leu Thr Asn Ile Cys Ile Lys Leu 485 49017780DNABacillus licheniformis DSM 13gene(1)..(780)putative protein - putative transcriptional regulator - BLi03995 17atg act gtt gag acg aag gac cac cag ctg ctc ggt tca gta aaa aat 48Met Thr Val Glu Thr Lys Asp His Gln Leu Leu Gly Ser Val Lys Asn1 5 10 15gcg ctt cgg ctg ctt caa tcg ttt acc tta gac aca ccg gaa aag aaa 96Ala Leu Arg Leu Leu Gln Ser Phe Thr Leu Asp Thr Pro Glu Lys Lys 20 25 30cta acc gaa ctc gcc tcg tcg ctt ggc ctc gga aaa agc acg gtc tcg 144Leu Thr Glu Leu Ala Ser Ser Leu Gly Leu Gly Lys Ser Thr Val Ser 35 40 45cgc ctt ttg tct aca ttg gaa agc gaa ggg ttt gtc atg aag gac ccg 192Arg Leu Leu Ser Thr Leu Glu Ser Glu Gly Phe Val Met Lys Asp Pro 50 55 60gaa tcc aga aga tac aag ctc ggc tta tcg gtc ctg cag tta aac acc 240Glu Ser Arg Arg Tyr Lys Leu Gly Leu Ser Val Leu Gln Leu Asn Thr65 70 75 80att gtc aac tcc act ctt gaa atc aac cgc gaa tcc ctg ccg gtc ctc 288Ile Val Asn Ser Thr Leu Glu Ile Asn Arg Glu Ser Leu Pro Val Leu 85 90 95aaa cgg ctg acc gat gag acc ggc gaa acg tcg cat atc gcg atg cgc 336Lys Arg Leu Thr Asp Glu Thr Gly Glu Thr Ser His Ile Ala Met Arg 100 105 110cgc ggt ctc agc gtc gtt tat ctc aat cgg acg gaa tgc ccc cgc ccg 384Arg Gly Leu Ser Val Val Tyr Leu Asn Arg Thr Glu Cys Pro Arg Pro 115 120 125gtc gaa ctt ctc tca tat atc ggg cgg caa aac ccg atg cac tgc aca 432Val Glu Leu Leu Ser Tyr Ile Gly Arg Gln Asn Pro Met His Cys Thr 130 135 140agc tca ggt aag gtg ctt ctc gcc ttt gat gaa gag ggc ata ttg gac 480Ser Ser Gly Lys Val Leu Leu Ala Phe Asp Glu Glu Gly Ile Leu Asp145 150 155 160gag tat gtg aag gaa ggc ttg aaa agc tgt acg aaa gaa acc att aca 528Glu Tyr Val Lys Glu Gly Leu Lys Ser Cys Thr Lys Glu Thr Ile Thr 165 170 175gac ggg gac aaa ttg agg gat gtg ctg aaa acc gtt aaa gaa aac gga 576Asp Gly Asp Lys Leu Arg Asp Val Leu Lys Thr Val Lys Glu Asn Gly 180 185 190ttt gac gtc agt tgc ggg gaa ttc ata gat ggt gtc aca tca atc tca 624Phe Asp Val Ser Cys Gly Glu Phe Ile Asp Gly Val Thr Ser Ile Ser 195 200 205gct ccc gtc aga aac cac gca ggt cga gtg ttg tac gcc gtc agt gtc 672Ala Pro Val Arg Asn His Ala Gly Arg Val Leu Tyr Ala Val Ser Val 210 215 220gta ggc cct gca aaa cgc atc aag gcc cgg cag gcg aaa atc att tac 720Val Gly Pro Ala Lys Arg Ile Lys Ala Arg Gln Ala Lys Ile Ile Tyr225 230 235 240caa gtg aaa caa gcg gcc cgg gat att tcc gaa agg atc ggg tac tgg 768Gln Val Lys Gln Ala Ala Arg Asp Ile Ser Glu Arg Ile Gly Tyr Trp 245 250 255aaa cga ata taa 780Lys Arg Ile18259PRTBacillus licheniformis DSM 13 18Met Thr Val Glu Thr Lys Asp His Gln Leu Leu Gly Ser Val Lys Asn1 5 10 15Ala Leu Arg Leu Leu Gln Ser Phe Thr Leu Asp Thr Pro Glu Lys Lys 20 25 30Leu Thr Glu Leu Ala Ser Ser Leu Gly Leu Gly Lys Ser Thr Val Ser 35 40 45Arg Leu Leu Ser Thr Leu Glu Ser Glu Gly Phe Val Met Lys Asp Pro 50 55 60Glu Ser Arg Arg Tyr Lys Leu Gly Leu Ser Val Leu Gln Leu Asn Thr65 70 75 80Ile Val Asn Ser Thr Leu Glu Ile Asn Arg Glu Ser Leu Pro Val Leu 85 90 95Lys Arg Leu Thr Asp Glu Thr Gly Glu Thr Ser His Ile Ala Met Arg 100 105 110Arg Gly Leu Ser Val Val Tyr Leu Asn Arg Thr Glu Cys Pro Arg Pro 115 120 125Val Glu Leu Leu Ser Tyr Ile Gly Arg Gln Asn Pro Met His Cys Thr 130 135 140Ser Ser Gly Lys Val Leu Leu Ala Phe Asp Glu Glu Gly Ile Leu Asp145 150 155 160Glu Tyr Val Lys Glu Gly Leu Lys Ser Cys Thr Lys Glu Thr Ile Thr 165 170 175Asp Gly Asp Lys Leu Arg Asp Val Leu Lys Thr Val Lys Glu Asn Gly 180 185 190Phe Asp Val Ser Cys Gly Glu Phe Ile Asp Gly Val Thr Ser Ile Ser 195 200 205Ala Pro Val Arg Asn His Ala Gly Arg Val Leu Tyr Ala Val Ser Val 210 215 220Val Gly Pro Ala Lys Arg Ile Lys Ala Arg Gln Ala Lys Ile Ile Tyr225 230 235 240Gln Val Lys Gln Ala Ala Arg Asp Ile Ser Glu Arg Ile Gly Tyr Trp 245 250 255Lys Arg Ile19951DNABacillus licheniformis DSM 13gene(1)..(951)conserved hypothetical protein - unknown function - BLi03996 19ttg tat gat ctt cac acg cac ttt ata ccg cgc gat gtg ctg ctt tgg 48Leu Tyr Asp Leu His Thr His Phe Ile Pro Arg Asp Val Leu Leu Trp1 5 10 15ctg gaa gac aat gaa aag aga gtc cac gct gtc cgc gaa aaa aag gct 96Leu Glu Asp Asn Glu Lys Arg Val His Ala Val Arg Glu Lys Lys Ala 20 25 30gga tgc caa gcg gag ttt ttg acg gtc aat cac aaa tgg ggc ttt gaa 144Gly Cys Gln Ala Glu Phe Leu Thr Val Asn His Lys Trp Gly Phe Glu 35 40 45tta aaa gag ctt ttc tac cag gag gct tta tat ttg gaa gcc cag cag 192Leu Lys Glu Leu Phe Tyr Gln Glu Ala Leu Tyr Leu Glu Ala Gln Gln 50 55 60gag gcc ggc gtc aca cac tca ttg gta tcg ccg gtt ccc cag ctt ttc 240Glu Ala Gly Val Thr His Ser Leu Val Ser Pro Val Pro Gln Leu Phe65 70 75 80ctt tac gat ctc gcc gaa gaa atc acg gac gaa tta tct ttt gta tac 288Leu Tyr Asp Leu Ala Glu Glu Ile Thr Asp Glu Leu Ser Phe Val Tyr 85 90 95aac aaa gcg ctt gcc gaa tgg gtt aaa aaa tac gac ggc cgc ctc tcg 336Asn Lys Ala Leu Ala Glu Trp Val Lys Lys Tyr Asp Gly Arg Leu Ser 100 105 110gga ctc ggc aca gtc ccg ctg aac agc cct tca aag gcg agc gag cgc 384Gly Leu Gly Thr Val Pro Leu Asn Ser Pro Ser Lys Ala Ser Glu Arg 115 120 125ctg aat gaa gcg atg aac aac ggg ctt aaa gga gcg att atc gga ccc 432Leu Asn Glu Ala Met Asn Asn Gly Leu Lys Gly Ala Ile Ile Gly Pro 130 135 140ggc gct tcg caa cta ctt ctc tcg gac gaa gca ttt aca ccg ttt tgg 480Gly Ala Ser Gln Leu Leu Leu Ser Asp Glu Ala Phe Thr Pro Phe Trp145 150 155 160gaa acg gct gat aga aga aaa gcg att atc ttt ata cat ccg ctc ctt 528Glu Thr Ala Asp Arg Arg Lys Ala Ile Ile Phe Ile His Pro Leu Leu 165 170 175tct gaa gat ccg cgg ctc cgc cgc agg atg ctg cca aac ttg atc ggc 576Ser Glu Asp Pro Arg Leu Arg Arg Arg Met Leu Pro Asn Leu Ile Gly 180 185 190gtt ccg tgg gaa acg acg att tgt gcc gca gac ctt ttg tta agc ggg 624Val Pro Trp Glu Thr Thr Ile Cys Ala Ala Asp Leu Leu Leu Ser Gly 195 200 205ttt aca gac cgg ttt cgc aat gtg aaa att ctt ttg gcc cac gga ggc 672Phe Thr Asp Arg Phe Arg Asn Val Lys Ile Leu Leu Ala His Gly Gly 210 215 220ggc ttt ttg cct tat cag atc gga aga ctg aac aaa ggg tat gac aaa 720Gly Phe Leu Pro Tyr Gln Ile Gly Arg Leu Asn Lys Gly Tyr Asp Lys225 230 235 240tgg gcc ggc gtt tct tca tcg ctt tcg gct ccg ccg cgc gaa tat ttg 768Trp Ala Gly Val Ser Ser Ser Leu Ser Ala Pro Pro Arg Glu Tyr Leu 245 250 255cgg aag ttt tgg ttt gat ggt gtg cta tgg gag caa gaa agc gcg gca 816Arg Lys Phe Trp Phe Asp Gly Val Leu Trp Glu Gln Glu Ser Ala Ala 260 265 270tat ttg aag aag ctt gtc gga gaa gac cgc ctt gtt ccg ggt tcc gat 864Tyr Leu Lys Lys Leu Val Gly Glu Asp Arg Leu Val Pro Gly Ser Asp 275 280 285ttt cct ttc gac ctt tgt cag tgg ccg cct ctc gat ccc ggc cgg aaa 912Phe Pro Phe Asp Leu Cys Gln Trp Pro Pro Leu Asp Pro Gly Arg Lys 290 295 300gga gcg gag tct ctg ctg aat gta gcg att gaa gca taa 951Gly Ala Glu Ser Leu Leu Asn Val Ala Ile Glu Ala305 310 31520316PRTBacillus licheniformis DSM 13misc_feature(1)..(1)Xaa can be Leu or Met. 20Xaa Tyr Asp Leu His Thr His Phe Ile Pro Arg Asp Val Leu Leu Trp1 5 10 15Leu Glu Asp Asn Glu Lys Arg Val His Ala Val Arg Glu Lys Lys Ala 20 25 30Gly Cys Gln Ala Glu Phe Leu Thr Val Asn His Lys Trp Gly Phe Glu 35 40 45Leu Lys Glu Leu Phe Tyr Gln Glu Ala Leu Tyr Leu Glu Ala Gln Gln 50 55 60Glu Ala Gly Val Thr His Ser Leu Val Ser Pro Val Pro Gln Leu Phe65 70 75 80Leu Tyr Asp Leu Ala Glu Glu Ile Thr Asp Glu Leu Ser Phe Val Tyr 85 90 95Asn Lys Ala Leu Ala Glu Trp Val Lys Lys Tyr Asp Gly Arg Leu Ser 100 105 110Gly Leu Gly Thr Val Pro Leu Asn Ser Pro Ser Lys Ala Ser Glu Arg 115 120 125Leu Asn Glu Ala Met Asn Asn Gly Leu Lys Gly Ala Ile Ile Gly Pro 130 135 140Gly Ala Ser Gln Leu Leu Leu Ser Asp Glu Ala Phe Thr Pro Phe Trp145 150 155 160Glu Thr Ala Asp Arg Arg Lys Ala Ile Ile Phe Ile His Pro Leu Leu 165 170 175Ser Glu Asp Pro Arg Leu Arg Arg Arg Met Leu Pro Asn Leu Ile Gly 180 185 190Val Pro Trp Glu Thr Thr Ile Cys Ala Ala Asp Leu Leu Leu Ser Gly 195 200 205Phe Thr Asp Arg Phe Arg Asn Val Lys Ile Leu Leu Ala His Gly Gly 210 215 220Gly Phe Leu Pro Tyr Gln Ile Gly Arg Leu Asn Lys Gly Tyr Asp Lys225 230 235 240Trp Ala Gly Val Ser Ser Ser Leu Ser Ala Pro Pro Arg Glu Tyr Leu 245 250 255Arg Lys Phe Trp Phe Asp Gly Val Leu Trp Glu Gln Glu Ser Ala Ala 260 265 270Tyr Leu Lys Lys Leu Val Gly Glu Asp Arg Leu Val Pro Gly Ser Asp 275 280 285Phe Pro Phe Asp Leu Cys Gln Trp Pro Pro Leu Asp Pro Gly Arg Lys 290 295 300Gly Ala Glu Ser Leu Leu Asn Val Ala Ile Glu Ala305 310 31521921DNABacillus licheniformis DSM 13gene(1)..(921)putative protein - putative phage capsid protein - BLi01470 21atg agt ttt gat ccg aat aac gta ttg atg caa gat gca gtc aaa gga 48Met Ser Phe Asp Pro Asn Asn Val Leu Met Gln Asp Ala Val Lys Gly1 5 10 15aaa gta cca tct gat caa gga aca tta gta tta aaa gac ttt atg aca 96Lys Val Pro Ser Asp Gln Gly Thr Leu Val Leu Lys Asp Phe Met Thr 20 25 30caa tca gca gtc aca aaa cta gca aaa tat gaa gaa atg gat aaa acc 144Gln Ser Ala Val Thr Lys Leu Ala Lys Tyr Glu Glu Met Asp Lys Thr 35 40 45gag aag aca ttc act tat ctt gcg tct ggc ccc ggg gct tac tgg gtt 192Glu Lys Thr Phe Thr Tyr Leu Ala Ser Gly Pro Gly Ala Tyr Trp Val 50 55 60ggc gaa ggt gag aga atc aaa act tct aag gct aca tgg tta gaa gcg 240Gly Glu Gly Glu Arg Ile Lys Thr Ser Lys Ala Thr Trp Leu Glu Ala65 70 75 80aaa atg acc tct aaa aaa ctc ggg gtc att gtc cca gtc aca aaa gag 288Lys Met Thr Ser Lys Lys Leu Gly Val Ile Val Pro Val Thr Lys Glu 85 90 95ttt ttg aat tac tcc gtg aaa gac ttc ttt act caa atg cgg cca gca 336Phe Leu Asn Tyr Ser Val Lys Asp Phe Phe Thr Gln Met Arg Pro Ala 100 105 110atc gcg gaa gca ttc gct att aaa ttt gac caa gca gca ctt ttt ggc 384Ile Ala Glu Ala Phe Ala Ile Lys Phe Asp Gln Ala Ala Leu Phe Gly 115 120 125att gat tca cca ttc gga aaa ggc act tct gta ttc gaa aga gca gaa 432Ile Asp Ser Pro Phe Gly Lys Gly Thr Ser Val Phe Glu Arg Ala Glu 130 135 140gca gca gga aac act gtt gtt ttg aac tct cta ggc aac ctt tac gat 480Ala Ala Gly Asn Thr Val Val Leu Asn Ser Leu Gly Asn Leu Tyr Asp145 150 155 160gag tta aac gca gtc atg gca ctc act gag gac aat gac aaa gac gtg 528Glu Leu Asn Ala Val Met Ala Leu Thr Glu Asp Asn Asp Lys Asp Val 165 170 175aac ggc ttt act act aca cgt cgt ttc aaa caa aaa cta cgc ggc act 576Asn Gly Phe Thr Thr Thr Arg Arg Phe Lys Gln Lys Leu Arg Gly Thr 180 185 190aag gac ggc aac ggg ctg cct att ttt aat gat gca aga ggc ggc gcg 624Lys Asp Gly Asn Gly Leu Pro Ile Phe Asn Asp Ala Arg Gly Gly Ala 195 200 205aca tca gag gca tta ggt ttg ccg att ggc tat gtc gat tct aag tct 672Thr Ser Glu Ala Leu Gly Leu Pro Ile Gly Tyr Val Asp Ser Lys Ser 210 215 220tgg gat tat aca aaa gca cat cta ttg gct gcc gat tgg gat ttt acc 720Trp Asp Tyr Thr Lys Ala His Leu Leu Ala Ala Asp Trp Asp Phe Thr225 230

235 240cgt tac gga att cct caa ggt atg gaa tac cat att tcg caa gat gca 768Arg Tyr Gly Ile Pro Gln Gly Met Glu Tyr His Ile Ser Gln Asp Ala 245 250 255act tta aca acg gtt gtc gat gaa aat aac aat cct atc aac ttg ttt 816Thr Leu Thr Thr Val Val Asp Glu Asn Asn Asn Pro Ile Asn Leu Phe 260 265 270gag cgt gat atg ttt gct ctc cga gtg act caa caa gtc gga ttt atg 864Glu Arg Asp Met Phe Ala Leu Arg Val Thr Gln Gln Val Gly Phe Met 275 280 285aca ctt tct gac gaa gca ttc gca gct ctt act ccg gaa gca gaa gcg 912Thr Leu Ser Asp Glu Ala Phe Ala Ala Leu Thr Pro Glu Ala Glu Ala 290 295 300ggg gcg taa 921Gly Ala30522306PRTBacillus licheniformis DSM 13 22Met Ser Phe Asp Pro Asn Asn Val Leu Met Gln Asp Ala Val Lys Gly1 5 10 15Lys Val Pro Ser Asp Gln Gly Thr Leu Val Leu Lys Asp Phe Met Thr 20 25 30Gln Ser Ala Val Thr Lys Leu Ala Lys Tyr Glu Glu Met Asp Lys Thr 35 40 45Glu Lys Thr Phe Thr Tyr Leu Ala Ser Gly Pro Gly Ala Tyr Trp Val 50 55 60Gly Glu Gly Glu Arg Ile Lys Thr Ser Lys Ala Thr Trp Leu Glu Ala65 70 75 80Lys Met Thr Ser Lys Lys Leu Gly Val Ile Val Pro Val Thr Lys Glu 85 90 95Phe Leu Asn Tyr Ser Val Lys Asp Phe Phe Thr Gln Met Arg Pro Ala 100 105 110Ile Ala Glu Ala Phe Ala Ile Lys Phe Asp Gln Ala Ala Leu Phe Gly 115 120 125Ile Asp Ser Pro Phe Gly Lys Gly Thr Ser Val Phe Glu Arg Ala Glu 130 135 140Ala Ala Gly Asn Thr Val Val Leu Asn Ser Leu Gly Asn Leu Tyr Asp145 150 155 160Glu Leu Asn Ala Val Met Ala Leu Thr Glu Asp Asn Asp Lys Asp Val 165 170 175Asn Gly Phe Thr Thr Thr Arg Arg Phe Lys Gln Lys Leu Arg Gly Thr 180 185 190Lys Asp Gly Asn Gly Leu Pro Ile Phe Asn Asp Ala Arg Gly Gly Ala 195 200 205Thr Ser Glu Ala Leu Gly Leu Pro Ile Gly Tyr Val Asp Ser Lys Ser 210 215 220Trp Asp Tyr Thr Lys Ala His Leu Leu Ala Ala Asp Trp Asp Phe Thr225 230 235 240Arg Tyr Gly Ile Pro Gln Gly Met Glu Tyr His Ile Ser Gln Asp Ala 245 250 255Thr Leu Thr Thr Val Val Asp Glu Asn Asn Asn Pro Ile Asn Leu Phe 260 265 270Glu Arg Asp Met Phe Ala Leu Arg Val Thr Gln Gln Val Gly Phe Met 275 280 285Thr Leu Ser Asp Glu Ala Phe Ala Ala Leu Thr Pro Glu Ala Glu Ala 290 295 300Gly Ala305231371DNABacillus licheniformis DSM 13gene(1)..(1371)yvtA - unknown function - similar to HtrA-like serine protease - BLi03481 23atg gat ttt aga cgc gat gac gaa caa aaa cat gca aat gaa gag cag 48Met Asp Phe Arg Arg Asp Asp Glu Gln Lys His Ala Asn Glu Glu Gln1 5 10 15ttt cac gaa gaa gcg cag acg ccg caa acg gca ggt tct gag gag cag 96Phe His Glu Glu Ala Gln Thr Pro Gln Thr Ala Gly Ser Glu Glu Gln 20 25 30cag tca gaa cag ccg gca aag gag ccc gag ctg att tta aag aag gat 144Gln Ser Glu Gln Pro Ala Lys Glu Pro Glu Leu Ile Leu Lys Lys Asp 35 40 45aca gac gat cag tca gat ctt acg aat aaa gaa acc gct gca gcc ctg 192Thr Asp Asp Gln Ser Asp Leu Thr Asn Lys Glu Thr Ala Ala Ala Leu 50 55 60gaa gat gcc agg gaa acg aga gcg gca aaa gaa aag cgg cgc aag gct 240Glu Asp Ala Arg Glu Thr Arg Ala Ala Lys Glu Lys Arg Arg Lys Ala65 70 75 80tcg tgg ctg agc ccg att ttg ggc ggg atc atc gga ggc ggc ttg gtg 288Ser Trp Leu Ser Pro Ile Leu Gly Gly Ile Ile Gly Gly Gly Leu Val 85 90 95ctt gga atc tcc ccg tat ctg ccg ggc gat cat ccg agc gaa act gcc 336Leu Gly Ile Ser Pro Tyr Leu Pro Gly Asp His Pro Ser Glu Thr Ala 100 105 110gga acc gga cag aca gag cca aag cag tcg cag aat ttc acc aca aaa 384Gly Thr Gly Gln Thr Glu Pro Lys Gln Ser Gln Asn Phe Thr Thr Lys 115 120 125cct gtc acg aac gca gat aac gta cct gat atg gtg gaa gat ttg gag 432Pro Val Thr Asn Ala Asp Asn Val Pro Asp Met Val Glu Asp Leu Glu 130 135 140cct gcg atc gtc gga gtt tca aat att caa acg agc ttc ggc ttc tct 480Pro Ala Ile Val Gly Val Ser Asn Ile Gln Thr Ser Phe Gly Phe Ser145 150 155 160gaa gac gac gtg gag gag agc ggc acg gga tca ggc gtc atc ttt aag 528Glu Asp Asp Val Glu Glu Ser Gly Thr Gly Ser Gly Val Ile Phe Lys 165 170 175aaa gac ggc ggc aag gcc tac att atc aca aac aac cat gtc gtt gaa 576Lys Asp Gly Gly Lys Ala Tyr Ile Ile Thr Asn Asn His Val Val Glu 180 185 190ggc gcc tcg aag gtg acc atc tcc tta tac aac gga aag acc gcg gat 624Gly Ala Ser Lys Val Thr Ile Ser Leu Tyr Asn Gly Lys Thr Ala Asp 195 200 205gcc aag att atc ggc agc gac gct ttg act gat ctg gct gtt tta gag 672Ala Lys Ile Ile Gly Ser Asp Ala Leu Thr Asp Leu Ala Val Leu Glu 210 215 220atc agc agc aaa ggc gtt gat aag gtt gca agt ttc gga gac tcc gca 720Ile Ser Ser Lys Gly Val Asp Lys Val Ala Ser Phe Gly Asp Ser Ala225 230 235 240aaa ctg cgc gcc ggc gaa aaa gta att gcg atc ggg aac cct ctc ggc 768Lys Leu Arg Ala Gly Glu Lys Val Ile Ala Ile Gly Asn Pro Leu Gly 245 250 255ctt caa ttt tca aga acg gtt aca gaa ggc atc atc agc ggt gtc aac 816Leu Gln Phe Ser Arg Thr Val Thr Glu Gly Ile Ile Ser Gly Val Asn 260 265 270cgg acg att gaa gtc tca aca tct gaa ggg aat tgg gat atg aac gtc 864Arg Thr Ile Glu Val Ser Thr Ser Glu Gly Asn Trp Asp Met Asn Val 275 280 285ctt cag aca gat gcg gcg atc aac cct gga aac agc ggc ggg cct ctg 912Leu Gln Thr Asp Ala Ala Ile Asn Pro Gly Asn Ser Gly Gly Pro Leu 290 295 300att aac agc agc ggg cag gtc atc ggc atc aac agc ctg aag atc agc 960Ile Asn Ser Ser Gly Gln Val Ile Gly Ile Asn Ser Leu Lys Ile Ser305 310 315 320caa agc ggt gtt gaa tcg ctt ggc ttc gcg att ccg agc aac gat gtt 1008Gln Ser Gly Val Glu Ser Leu Gly Phe Ala Ile Pro Ser Asn Asp Val 325 330 335cag ccg atc gtt gac gag ctt ttg gaa aaa ggg aaa gtg gaa cgg cct 1056Gln Pro Ile Val Asp Glu Leu Leu Glu Lys Gly Lys Val Glu Arg Pro 340 345 350ttc ctc ggc gtg caa atg atc gat atg cag cag gtg cct gaa cag tac 1104Phe Leu Gly Val Gln Met Ile Asp Met Gln Gln Val Pro Glu Gln Tyr 355 360 365cag caa aat acg ctc ggc cta ttc gga gac cag ctt aat aaa ggc gtg 1152Gln Gln Asn Thr Leu Gly Leu Phe Gly Asp Gln Leu Asn Lys Gly Val 370 375 380tac atc gat aaa gta tcg ccg aaa tcg cct gcg gca gac gcg ggt atg 1200Tyr Ile Asp Lys Val Ser Pro Lys Ser Pro Ala Ala Asp Ala Gly Met385 390 395 400aaa gcc ggc gac gtc atc acc aaa atg aac ggc aaa aac gtc gaa acc 1248Lys Ala Gly Asp Val Ile Thr Lys Met Asn Gly Lys Asn Val Glu Thr 405 410 415acc tca gac ctc aga aaa att ctt tat aca gaa gca aaa gca gga gat 1296Thr Ser Asp Leu Arg Lys Ile Leu Tyr Thr Glu Ala Lys Ala Gly Asp 420 425 430act gtc aca ttt gag gtg ctg aga aac gga aag caa aca aca atg aaa 1344Thr Val Thr Phe Glu Val Leu Arg Asn Gly Lys Gln Thr Thr Met Lys 435 440 445gca aaa ctt gcg aaa agc aaa tca taa 1371Ala Lys Leu Ala Lys Ser Lys Ser 450 45524456PRTBacillus licheniformis DSM 13 24Met Asp Phe Arg Arg Asp Asp Glu Gln Lys His Ala Asn Glu Glu Gln1 5 10 15Phe His Glu Glu Ala Gln Thr Pro Gln Thr Ala Gly Ser Glu Glu Gln 20 25 30Gln Ser Glu Gln Pro Ala Lys Glu Pro Glu Leu Ile Leu Lys Lys Asp 35 40 45Thr Asp Asp Gln Ser Asp Leu Thr Asn Lys Glu Thr Ala Ala Ala Leu 50 55 60Glu Asp Ala Arg Glu Thr Arg Ala Ala Lys Glu Lys Arg Arg Lys Ala65 70 75 80Ser Trp Leu Ser Pro Ile Leu Gly Gly Ile Ile Gly Gly Gly Leu Val 85 90 95Leu Gly Ile Ser Pro Tyr Leu Pro Gly Asp His Pro Ser Glu Thr Ala 100 105 110Gly Thr Gly Gln Thr Glu Pro Lys Gln Ser Gln Asn Phe Thr Thr Lys 115 120 125Pro Val Thr Asn Ala Asp Asn Val Pro Asp Met Val Glu Asp Leu Glu 130 135 140Pro Ala Ile Val Gly Val Ser Asn Ile Gln Thr Ser Phe Gly Phe Ser145 150 155 160Glu Asp Asp Val Glu Glu Ser Gly Thr Gly Ser Gly Val Ile Phe Lys 165 170 175Lys Asp Gly Gly Lys Ala Tyr Ile Ile Thr Asn Asn His Val Val Glu 180 185 190Gly Ala Ser Lys Val Thr Ile Ser Leu Tyr Asn Gly Lys Thr Ala Asp 195 200 205Ala Lys Ile Ile Gly Ser Asp Ala Leu Thr Asp Leu Ala Val Leu Glu 210 215 220Ile Ser Ser Lys Gly Val Asp Lys Val Ala Ser Phe Gly Asp Ser Ala225 230 235 240Lys Leu Arg Ala Gly Glu Lys Val Ile Ala Ile Gly Asn Pro Leu Gly 245 250 255Leu Gln Phe Ser Arg Thr Val Thr Glu Gly Ile Ile Ser Gly Val Asn 260 265 270Arg Thr Ile Glu Val Ser Thr Ser Glu Gly Asn Trp Asp Met Asn Val 275 280 285Leu Gln Thr Asp Ala Ala Ile Asn Pro Gly Asn Ser Gly Gly Pro Leu 290 295 300Ile Asn Ser Ser Gly Gln Val Ile Gly Ile Asn Ser Leu Lys Ile Ser305 310 315 320Gln Ser Gly Val Glu Ser Leu Gly Phe Ala Ile Pro Ser Asn Asp Val 325 330 335Gln Pro Ile Val Asp Glu Leu Leu Glu Lys Gly Lys Val Glu Arg Pro 340 345 350Phe Leu Gly Val Gln Met Ile Asp Met Gln Gln Val Pro Glu Gln Tyr 355 360 365Gln Gln Asn Thr Leu Gly Leu Phe Gly Asp Gln Leu Asn Lys Gly Val 370 375 380Tyr Ile Asp Lys Val Ser Pro Lys Ser Pro Ala Ala Asp Ala Gly Met385 390 395 400Lys Ala Gly Asp Val Ile Thr Lys Met Asn Gly Lys Asn Val Glu Thr 405 410 415Thr Ser Asp Leu Arg Lys Ile Leu Tyr Thr Glu Ala Lys Ala Gly Asp 420 425 430Thr Val Thr Phe Glu Val Leu Arg Asn Gly Lys Gln Thr Thr Met Lys 435 440 445Ala Lys Leu Ala Lys Ser Lys Ser 450 455251017DNABacillus licheniformis DSM 13gene(1)..(1017)kdgR - transcriptional repressor of the pectin utilization operon - BLi03828 25atg aag gat aag caa aca gcg aaa gta acc atc aac gaa gtc gcc gca 48Met Lys Asp Lys Gln Thr Ala Lys Val Thr Ile Asn Glu Val Ala Ala1 5 10 15cat gca ggc gta tca aaa tca acg gtt tcc cgt tat atc aac ggg aag 96His Ala Gly Val Ser Lys Ser Thr Val Ser Arg Tyr Ile Asn Gly Lys 20 25 30att gat gcc att tca ccg aaa aag gtg aaa agc atc aaa aaa gcg atc 144Ile Asp Ala Ile Ser Pro Lys Lys Val Lys Ser Ile Lys Lys Ala Ile 35 40 45gag gag ctc aat tac cgc cca agc cag atg gca cag ggg ctg aag gtg 192Glu Glu Leu Asn Tyr Arg Pro Ser Gln Met Ala Gln Gly Leu Lys Val 50 55 60aaa aaa agc aaa gtt atc gga ttt ttg gtc gcc gat att aca aac ccg 240Lys Lys Ser Lys Val Ile Gly Phe Leu Val Ala Asp Ile Thr Asn Pro65 70 75 80ttt tca gta gcc acc ttg aga ggg gtg gaa gag gtt tgc gat cag tac 288Phe Ser Val Ala Thr Leu Arg Gly Val Glu Glu Val Cys Asp Gln Tyr 85 90 95gga tac agc atc atg gtc tgc aac acg gac aac agt cct gag aaa gaa 336Gly Tyr Ser Ile Met Val Cys Asn Thr Asp Asn Ser Pro Glu Lys Glu 100 105 110agg gag atg ctg cac aag ctg aac gcg cat tac att gaa ggg ctg att 384Arg Glu Met Leu His Lys Leu Asn Ala His Tyr Ile Glu Gly Leu Ile 115 120 125att aat acg acg gga agg aac aac gat gtg ctc ttg gat tta atc agc 432Ile Asn Thr Thr Gly Arg Asn Asn Asp Val Leu Leu Asp Leu Ile Ser 130 135 140cag gat gta tcg att gtg ctc gtt gac cgg aaa gtg ccc ggg ctc aag 480Gln Asp Val Ser Ile Val Leu Val Asp Arg Lys Val Pro Gly Leu Lys145 150 155 160atc gat gca gtc aca acg aac aac cgc gaa gtg aca agt caa atc gtc 528Ile Asp Ala Val Thr Thr Asn Asn Arg Glu Val Thr Ser Gln Ile Val 165 170 175aat ctc atg tat gac aaa ggc tat gcc cag gtc ggt ctg ttt acc gaa 576Asn Leu Met Tyr Asp Lys Gly Tyr Ala Gln Val Gly Leu Phe Thr Glu 180 185 190ccg att aag gga atc agt cca agg gag gaa agg gcc tcc gcc tat aca 624Pro Ile Lys Gly Ile Ser Pro Arg Glu Glu Arg Ala Ser Ala Tyr Thr 195 200 205gaa atc gcg ctg gaa cga aac gcc ggc ggt acc cct ctc gta tat gag 672Glu Ile Ala Leu Glu Arg Asn Ala Gly Gly Thr Pro Leu Val Tyr Glu 210 215 220gct gac gtc aaa gac aaa gaa tct cta tta cag tcc gtc aaa agc ttt 720Ala Asp Val Lys Asp Lys Glu Ser Leu Leu Gln Ser Val Lys Ser Phe225 230 235 240ttg gaa atg aaa gaa ggg aaa aaa gcc att ctt gga aac aac ggg ctt 768Leu Glu Met Lys Glu Gly Lys Lys Ala Ile Leu Gly Asn Asn Gly Leu 245 250 255ctg atg ctg aaa atc atc agc tgc ctt tat gag ctt ggc att tca att 816Leu Met Leu Lys Ile Ile Ser Cys Leu Tyr Glu Leu Gly Ile Ser Ile 260 265 270ccg gaa gat gtc gga att gcc gga ttt gat gat aca gaa tgg tat aaa 864Pro Glu Asp Val Gly Ile Ala Gly Phe Asp Asp Thr Glu Trp Tyr Lys 275 280 285ttg atc ggt ccg ggc att acg acc atc gcc cag ccg tcg cat gaa atg 912Leu Ile Gly Pro Gly Ile Thr Thr Ile Ala Gln Pro Ser His Glu Met 290 295 300ggc agg gtc gcc atg gag cgg atc atc aaa cgg att gaa gga gac gaa 960Gly Arg Val Ala Met Glu Arg Ile Ile Lys Arg Ile Glu Gly Asp Glu305 310 315 320agc gcc ccg cag acg att cag ctt gag gcg gag ctt gtt ctc aga cag 1008Ser Ala Pro Gln Thr Ile Gln Leu Glu Ala Glu Leu Val Leu Arg Gln 325 330 335tcg ctg tag 1017Ser Leu26338PRTBacillus licheniformis DSM 13 26Met Lys Asp Lys Gln Thr Ala Lys Val Thr Ile Asn Glu Val Ala Ala1 5 10 15His Ala Gly Val Ser Lys Ser Thr Val Ser Arg Tyr Ile Asn Gly Lys 20 25 30Ile Asp Ala Ile Ser Pro Lys Lys Val Lys Ser Ile Lys Lys Ala Ile 35 40 45Glu Glu Leu Asn Tyr Arg Pro Ser Gln Met Ala Gln Gly Leu Lys Val 50 55 60Lys Lys Ser Lys Val Ile Gly Phe Leu Val Ala Asp Ile Thr Asn Pro65 70 75 80Phe Ser Val Ala Thr Leu Arg Gly Val Glu Glu Val Cys Asp Gln Tyr 85 90 95Gly Tyr Ser Ile Met Val Cys Asn Thr Asp Asn Ser Pro Glu Lys Glu 100 105 110Arg Glu Met Leu His Lys Leu Asn Ala His Tyr Ile Glu Gly Leu Ile 115 120 125Ile Asn Thr Thr Gly Arg Asn Asn Asp Val Leu Leu Asp Leu Ile Ser 130 135 140Gln Asp Val Ser Ile Val Leu Val Asp Arg Lys Val Pro Gly Leu Lys145 150 155 160Ile Asp Ala Val Thr Thr Asn Asn Arg Glu Val Thr Ser Gln Ile Val 165 170 175Asn Leu Met Tyr Asp Lys Gly Tyr Ala Gln Val Gly Leu Phe Thr Glu 180 185 190Pro Ile Lys Gly Ile Ser Pro Arg Glu Glu Arg Ala Ser Ala Tyr Thr 195 200 205Glu Ile Ala Leu Glu Arg Asn Ala Gly Gly Thr Pro Leu Val Tyr Glu 210 215 220Ala Asp Val Lys Asp Lys Glu Ser Leu Leu Gln Ser Val Lys Ser Phe225 230 235 240Leu Glu Met Lys Glu Gly Lys Lys Ala Ile Leu Gly Asn Asn Gly Leu 245 250 255Leu Met Leu

Lys Ile Ile Ser Cys Leu Tyr Glu Leu Gly Ile Ser Ile 260 265 270Pro Glu Asp Val Gly Ile Ala Gly Phe Asp Asp Thr Glu Trp Tyr Lys 275 280 285Leu Ile Gly Pro Gly Ile Thr Thr Ile Ala Gln Pro Ser His Glu Met 290 295 300Gly Arg Val Ala Met Glu Arg Ile Ile Lys Arg Ile Glu Gly Asp Glu305 310 315 320Ser Ala Pro Gln Thr Ile Gln Leu Glu Ala Glu Leu Val Leu Arg Gln 325 330 335Ser Leu27645DNABacillus licheniformis DSM 13gene(1)..(645)ycnI - unknown function - similar to unknown function proteins - BLi00479 27atg ttg aaa aag aca tta tgg atg ggt gca atc cta gca tca ttt ttg 48Met Leu Lys Lys Thr Leu Trp Met Gly Ala Ile Leu Ala Ser Phe Leu1 5 10 15ctt ttc tcc gtg ccg gtc agc gca cat gtc acg gta aaa ccg acc gaa 96Leu Phe Ser Val Pro Val Ser Ala His Val Thr Val Lys Pro Thr Glu 20 25 30tca gca gca ggc tca tgg gaa aca tac act gtc aaa gta ccg gtt gaa 144Ser Ala Ala Gly Ser Trp Glu Thr Tyr Thr Val Lys Val Pro Val Glu 35 40 45aaa gac aac gcc acg atc aag gtc gtt ctc agc atg ccg gac ggt gtt 192Lys Asp Asn Ala Thr Ile Lys Val Val Leu Ser Met Pro Asp Gly Val 50 55 60gaa ttt cag cag tat gaa ccg gtg ccg ggg tgg aaa aca agc act gaa 240Glu Phe Gln Gln Tyr Glu Pro Val Pro Gly Trp Lys Thr Ser Thr Glu65 70 75 80acc ggc aaa aac gaa aaa gta aca agg gtg acg tgg gaa gcc gaa aac 288Thr Gly Lys Asn Glu Lys Val Thr Arg Val Thr Trp Glu Ala Glu Asn 85 90 95ggc ggc att cag cct ggt gaa ttc cag cag ttt acc ttc acc gcc aaa 336Gly Gly Ile Gln Pro Gly Glu Phe Gln Gln Phe Thr Phe Thr Ala Lys 100 105 110aac cct gac aaa gaa caa aag gcc gcc tgg aac gca tac caa tat tat 384Asn Pro Asp Lys Glu Gln Lys Ala Ala Trp Asn Ala Tyr Gln Tyr Tyr 115 120 125aag gac ggc aca att gtc gag tgg acc ggt gat gat gac gcc gat acc 432Lys Asp Gly Thr Ile Val Glu Trp Thr Gly Asp Asp Asp Ala Asp Thr 130 135 140ccg cat gca atc aca agc att gtc gac gct gca aaa ctc gga cac gaa 480Pro His Ala Ile Thr Ser Ile Val Asp Ala Ala Lys Leu Gly His Glu145 150 155 160acg gtt gat gca cac ggc gct tca acg aaa caa aac gct gcc gca gaa 528Thr Val Asp Ala His Gly Ala Ser Thr Lys Gln Asn Ala Ala Ala Glu 165 170 175aat gaa gac agt caa gga gga caa ggg att caa acc gct tcc ctg atc 576Asn Glu Asp Ser Gln Gly Gly Gln Gly Ile Gln Thr Ala Ser Leu Ile 180 185 190ctg tcg atc ctc gcc gtg ctt ctt gga ggt gct gcg ctg ctc ttg aca 624Leu Ser Ile Leu Ala Val Leu Leu Gly Gly Ala Ala Leu Leu Leu Thr 195 200 205gtg cgg aaa aag aaa tcg taa 645Val Arg Lys Lys Lys Ser 21028214PRTBacillus licheniformis DSM 13 28Met Leu Lys Lys Thr Leu Trp Met Gly Ala Ile Leu Ala Ser Phe Leu1 5 10 15Leu Phe Ser Val Pro Val Ser Ala His Val Thr Val Lys Pro Thr Glu 20 25 30Ser Ala Ala Gly Ser Trp Glu Thr Tyr Thr Val Lys Val Pro Val Glu 35 40 45Lys Asp Asn Ala Thr Ile Lys Val Val Leu Ser Met Pro Asp Gly Val 50 55 60Glu Phe Gln Gln Tyr Glu Pro Val Pro Gly Trp Lys Thr Ser Thr Glu65 70 75 80Thr Gly Lys Asn Glu Lys Val Thr Arg Val Thr Trp Glu Ala Glu Asn 85 90 95Gly Gly Ile Gln Pro Gly Glu Phe Gln Gln Phe Thr Phe Thr Ala Lys 100 105 110Asn Pro Asp Lys Glu Gln Lys Ala Ala Trp Asn Ala Tyr Gln Tyr Tyr 115 120 125Lys Asp Gly Thr Ile Val Glu Trp Thr Gly Asp Asp Asp Ala Asp Thr 130 135 140Pro His Ala Ile Thr Ser Ile Val Asp Ala Ala Lys Leu Gly His Glu145 150 155 160Thr Val Asp Ala His Gly Ala Ser Thr Lys Gln Asn Ala Ala Ala Glu 165 170 175Asn Glu Asp Ser Gln Gly Gly Gln Gly Ile Gln Thr Ala Ser Leu Ile 180 185 190Leu Ser Ile Leu Ala Val Leu Leu Gly Gly Ala Ala Leu Leu Leu Thr 195 200 205Val Arg Lys Lys Lys Ser 210291647DNABacillus licheniformis DSM 13gene(1)..(1647)ycnJ - unknown function - similar to copper export protein - BLi00480 29atg aat gtg atg aaa aga gca aca ctc tgg gca gtt ctg gct gtt ctt 48Met Asn Val Met Lys Arg Ala Thr Leu Trp Ala Val Leu Ala Val Leu1 5 10 15tgt acc att ctg ctg cca aaa gcc gct ttt gct cac gct tat atc gta 96Cys Thr Ile Leu Leu Pro Lys Ala Ala Phe Ala His Ala Tyr Ile Val 20 25 30caa tca act cct gct gaa aat cag gag ctg aaa aaa gcg cct gaa gaa 144Gln Ser Thr Pro Ala Glu Asn Gln Glu Leu Lys Lys Ala Pro Glu Glu 35 40 45att aag atc gaa ttc aat gaa gcg att cag aac ggg ttt cat tcg ata 192Ile Lys Ile Glu Phe Asn Glu Ala Ile Gln Asn Gly Phe His Ser Ile 50 55 60acg gtc aga aat tcg tcg ggt gaa cgg gta gat tcc ggc aaa acg gaa 240Thr Val Arg Asn Ser Ser Gly Glu Arg Val Asp Ser Gly Lys Thr Glu65 70 75 80atc gac ccc caa aac gca aag atc atg acc gtc aag ctg aaa ccg aag 288Ile Asp Pro Gln Asn Ala Lys Ile Met Thr Val Lys Leu Lys Pro Lys 85 90 95tta aag gat gat atc tac tcg gcc gaa tgg aga gcc gta tcc gca gac 336Leu Lys Asp Asp Ile Tyr Ser Ala Glu Trp Arg Ala Val Ser Ala Asp 100 105 110ggt cat cct gta tca ggc acg atc cct ttt agt atc gga aac att caa 384Gly His Pro Val Ser Gly Thr Ile Pro Phe Ser Ile Gly Asn Ile Gln 115 120 125ggc ggg ctg ccg aaa gat cag caa aat caa gca tca ggc ctg ccg gca 432Gly Gly Leu Pro Lys Asp Gln Gln Asn Gln Ala Ser Gly Leu Pro Ala 130 135 140gcc gat atc gtc att gaa cgc gtc ctt ttg tat acc ggt ttc tcc ctg 480Ala Asp Ile Val Ile Glu Arg Val Leu Leu Tyr Thr Gly Phe Ser Leu145 150 155 160ttc atg gga gcc gct ctt ttt cag ctc gtc tgg cac cga cct gaa aac 528Phe Met Gly Ala Ala Leu Phe Gln Leu Val Trp His Arg Pro Glu Asn 165 170 175ggc tgg ccg ggt gac aaa aag cgc cgt ctg aag cgg atc acg aat gtg 576Gly Trp Pro Gly Asp Lys Lys Arg Arg Leu Lys Arg Ile Thr Asn Val 180 185 190gcg ctc tta ttg aca ggg gct gcc atc atc att cag ctg cca ctg caa 624Ala Leu Leu Leu Thr Gly Ala Ala Ile Ile Ile Gln Leu Pro Leu Gln 195 200 205aca aaa aca aac gca ggc gtt tcc tgg atc ggg gct ttt cag cct tcg 672Thr Lys Thr Asn Ala Gly Val Ser Trp Ile Gly Ala Phe Gln Pro Ser 210 215 220ctt tta aaa gaa acg ctt ctg aat acg acg ggc gga acg ctg tgg ctg 720Leu Leu Lys Glu Thr Leu Leu Asn Thr Thr Gly Gly Thr Leu Trp Leu225 230 235 240atc aac atc gcc atc atc gcc ttg ctt gtt ttc tct tcg aat aga aaa 768Ile Asn Ile Ala Ile Ile Ala Leu Leu Val Phe Ser Ser Asn Arg Lys 245 250 255ggc ctg aag agc gcg atc ttg agc ctg ttc ctt ttt gcc gga cta ttg 816Gly Leu Lys Ser Ala Ile Leu Ser Leu Phe Leu Phe Ala Gly Leu Leu 260 265 270ttc gtc aaa tcg ctc gac ggc cat gcc gct tcg acg agc gca ggc tat 864Phe Val Lys Ser Leu Asp Gly His Ala Ala Ser Thr Ser Ala Gly Tyr 275 280 285atg aca atc gcg atg aac ttc atc cat ctg aat gcc gct tcc att tgg 912Met Thr Ile Ala Met Asn Phe Ile His Leu Asn Ala Ala Ser Ile Trp 290 295 300act ggc gga ctc gcg atg ttt gtg ctg ttt ttc gga aaa gac tgg ctg 960Thr Gly Gly Leu Ala Met Phe Val Leu Phe Phe Gly Lys Asp Trp Leu305 310 315 320aag cct gac aaa agt ttg ata tgg gaa acc att cgg cgc ttc tcg cca 1008Lys Pro Asp Lys Ser Leu Ile Trp Glu Thr Ile Arg Arg Phe Ser Pro 325 330 335tgg gcg ctc gct tca gca ggg cta ctc ata ttc tcg ggc ctt ttc aac 1056Trp Ala Leu Ala Ser Ala Gly Leu Leu Ile Phe Ser Gly Leu Phe Asn 340 345 350agc ttt ttc att gtt cat aca ctg gat aac ctg ttc ggg acg gct tac 1104Ser Phe Phe Ile Val His Thr Leu Asp Asn Leu Phe Gly Thr Ala Tyr 355 360 365gga aaa gcg ctc ctt gtg aaa tcg gcg ctc gtc ctt ctc atg atc gga 1152Gly Lys Ala Leu Leu Val Lys Ser Ala Leu Val Leu Leu Met Ile Gly 370 375 380ctc ggg gca ttt cac tat ttt cgt ttt aga aaa gcg ccg aaa cag cag 1200Leu Gly Ala Phe His Tyr Phe Arg Phe Arg Lys Ala Pro Lys Gln Gln385 390 395 400ggc ggc cgt tcg atg aaa gct gaa tgg gcg gtc ggg ctt ctc att ctg 1248Gly Gly Arg Ser Met Lys Ala Glu Trp Ala Val Gly Leu Leu Ile Leu 405 410 415ctg acc acg gcg gtt ttc acc aat att cca agt ccg ccc gcg ccg ctg 1296Leu Thr Thr Ala Val Phe Thr Asn Ile Pro Ser Pro Pro Ala Pro Leu 420 425 430ccg cag cct ttc agt gag acg aag cag ctg acg gaa gga aat acg ctg 1344Pro Gln Pro Phe Ser Glu Thr Lys Gln Leu Thr Glu Gly Asn Thr Leu 435 440 445tca ctc cga atc agc cca aac atg ccg ggc tca aat acg ttt gaa gtc 1392Ser Leu Arg Ile Ser Pro Asn Met Pro Gly Ser Asn Thr Phe Glu Val 450 455 460acc gtc aga aac aga aaa gga gac gtc gtc aaa gat atc caa aaa att 1440Thr Val Arg Asn Arg Lys Gly Asp Val Val Lys Asp Ile Gln Lys Ile465 470 475 480gaa tta acc gtt gag aaa acc ggg ctt ttc ggg gat gac aag gaa agc 1488Glu Leu Thr Val Glu Lys Thr Gly Leu Phe Gly Asp Asp Lys Glu Ser 485 490 495acg ttc agc ctc agc aaa aat aaa gac ggt gtt ttc caa acg aaa aat 1536Thr Phe Ser Leu Ser Lys Asn Lys Asp Gly Val Phe Gln Thr Lys Asn 500 505 510tta aat tta agc gaa gac ggc atc tgg aag atc cgc gtt cac gga tta 1584Leu Asn Leu Ser Glu Asp Gly Ile Trp Lys Ile Arg Val His Gly Leu 515 520 525acg gcg tca ttt gag gaa atc aga aca gtt ttc aat gca aaa att aat 1632Thr Ala Ser Phe Glu Glu Ile Arg Thr Val Phe Asn Ala Lys Ile Asn 530 535 540aaa aag gag ttt tga 1647Lys Lys Glu Phe54530548PRTBacillus licheniformis DSM 13 30Met Asn Val Met Lys Arg Ala Thr Leu Trp Ala Val Leu Ala Val Leu1 5 10 15Cys Thr Ile Leu Leu Pro Lys Ala Ala Phe Ala His Ala Tyr Ile Val 20 25 30Gln Ser Thr Pro Ala Glu Asn Gln Glu Leu Lys Lys Ala Pro Glu Glu 35 40 45Ile Lys Ile Glu Phe Asn Glu Ala Ile Gln Asn Gly Phe His Ser Ile 50 55 60Thr Val Arg Asn Ser Ser Gly Glu Arg Val Asp Ser Gly Lys Thr Glu65 70 75 80Ile Asp Pro Gln Asn Ala Lys Ile Met Thr Val Lys Leu Lys Pro Lys 85 90 95Leu Lys Asp Asp Ile Tyr Ser Ala Glu Trp Arg Ala Val Ser Ala Asp 100 105 110Gly His Pro Val Ser Gly Thr Ile Pro Phe Ser Ile Gly Asn Ile Gln 115 120 125Gly Gly Leu Pro Lys Asp Gln Gln Asn Gln Ala Ser Gly Leu Pro Ala 130 135 140Ala Asp Ile Val Ile Glu Arg Val Leu Leu Tyr Thr Gly Phe Ser Leu145 150 155 160Phe Met Gly Ala Ala Leu Phe Gln Leu Val Trp His Arg Pro Glu Asn 165 170 175Gly Trp Pro Gly Asp Lys Lys Arg Arg Leu Lys Arg Ile Thr Asn Val 180 185 190Ala Leu Leu Leu Thr Gly Ala Ala Ile Ile Ile Gln Leu Pro Leu Gln 195 200 205Thr Lys Thr Asn Ala Gly Val Ser Trp Ile Gly Ala Phe Gln Pro Ser 210 215 220Leu Leu Lys Glu Thr Leu Leu Asn Thr Thr Gly Gly Thr Leu Trp Leu225 230 235 240Ile Asn Ile Ala Ile Ile Ala Leu Leu Val Phe Ser Ser Asn Arg Lys 245 250 255Gly Leu Lys Ser Ala Ile Leu Ser Leu Phe Leu Phe Ala Gly Leu Leu 260 265 270Phe Val Lys Ser Leu Asp Gly His Ala Ala Ser Thr Ser Ala Gly Tyr 275 280 285Met Thr Ile Ala Met Asn Phe Ile His Leu Asn Ala Ala Ser Ile Trp 290 295 300Thr Gly Gly Leu Ala Met Phe Val Leu Phe Phe Gly Lys Asp Trp Leu305 310 315 320Lys Pro Asp Lys Ser Leu Ile Trp Glu Thr Ile Arg Arg Phe Ser Pro 325 330 335Trp Ala Leu Ala Ser Ala Gly Leu Leu Ile Phe Ser Gly Leu Phe Asn 340 345 350Ser Phe Phe Ile Val His Thr Leu Asp Asn Leu Phe Gly Thr Ala Tyr 355 360 365Gly Lys Ala Leu Leu Val Lys Ser Ala Leu Val Leu Leu Met Ile Gly 370 375 380Leu Gly Ala Phe His Tyr Phe Arg Phe Arg Lys Ala Pro Lys Gln Gln385 390 395 400Gly Gly Arg Ser Met Lys Ala Glu Trp Ala Val Gly Leu Leu Ile Leu 405 410 415Leu Thr Thr Ala Val Phe Thr Asn Ile Pro Ser Pro Pro Ala Pro Leu 420 425 430Pro Gln Pro Phe Ser Glu Thr Lys Gln Leu Thr Glu Gly Asn Thr Leu 435 440 445Ser Leu Arg Ile Ser Pro Asn Met Pro Gly Ser Asn Thr Phe Glu Val 450 455 460Thr Val Arg Asn Arg Lys Gly Asp Val Val Lys Asp Ile Gln Lys Ile465 470 475 480Glu Leu Thr Val Glu Lys Thr Gly Leu Phe Gly Asp Asp Lys Glu Ser 485 490 495Thr Phe Ser Leu Ser Lys Asn Lys Asp Gly Val Phe Gln Thr Lys Asn 500 505 510Leu Asn Leu Ser Glu Asp Gly Ile Trp Lys Ile Arg Val His Gly Leu 515 520 525Thr Ala Ser Phe Glu Glu Ile Arg Thr Val Phe Asn Ala Lys Ile Asn 530 535 540Lys Lys Glu Phe54531573DNABacillus licheniformis DSM 13gene(1)..(573)ycnK - unknown function - similar to transcriptional regulator (DeoR family) - BLi00481 31atg cta ccg att aaa aga agg gag cag att ttg gct tgg atc aaa gaa 48Met Leu Pro Ile Lys Arg Arg Glu Gln Ile Leu Ala Trp Ile Lys Glu1 5 10 15gaa gaa tct ctg cgc att tcg gaa att tcg aaa agg ctg aac gtc tct 96Glu Glu Ser Leu Arg Ile Ser Glu Ile Ser Lys Arg Leu Asn Val Ser 20 25 30gaa atg acc gtt tac cgc gac att aaa ccg ctc gtt gaa aac ggg cag 144Glu Met Thr Val Tyr Arg Asp Ile Lys Pro Leu Val Glu Asn Gly Gln 35 40 45gtc atc aag acc gca ggc gga att acg cta aac aag ccg aag cag cag 192Val Ile Lys Thr Ala Gly Gly Ile Thr Leu Asn Lys Pro Lys Gln Gln 50 55 60ctt ggg cag ctt tgc gtc gtt tgc gga aaa ggg gca ggt tcg agg ctt 240Leu Gly Gln Leu Cys Val Val Cys Gly Lys Gly Ala Gly Ser Arg Leu65 70 75 80gcc gtg caa att gtc aaa aac gac agc caa att gaa cag ttt tgc tgc 288Ala Val Gln Ile Val Lys Asn Asp Ser Gln Ile Glu Gln Phe Cys Cys 85 90 95gtg cat tgc gca atg ctc cgc tat gag aaa gtg aaa gat gat gta tca 336Val His Cys Ala Met Leu Arg Tyr Glu Lys Val Lys Asp Asp Val Ser 100 105 110cag att atc tgc agg gat ttt ttg ctg gat acg acg atc agc gcc aaa 384Gln Ile Ile Cys Arg Asp Phe Leu Leu Asp Thr Thr Ile Ser Ala Lys 115 120 125gct gcc gtc ttt ttg ctg gat aca gat ctt cat ttg aac tgc tgc gag 432Ala Ala Val Phe Leu Leu Asp Thr Asp Leu His Leu Asn Cys Cys Glu 130 135 140ccg cat gcg ctc cct ttc gct tct gaa gca gaa gcg cgg aaa ttc aaa 480Pro His Ala Leu Pro Phe Ala Ser Glu Ala Glu Ala Arg Lys Phe Lys145 150 155 160aaa ggc ttt ggc gga cag ctg tta tcg ttt gac gat gcc gcg ctt ctg 528Lys Gly Phe Gly Gly Gln Leu Leu Ser Phe Asp Asp Ala Ala Leu Leu 165 170 175gtc cag aaa acg atg aag aac agc tgc tgc agc ctg aag aca tga 573Val Gln Lys Thr Met Lys Asn Ser Cys Cys Ser Leu Lys Thr 180 185 19032190PRTBacillus licheniformis DSM 13 32Met Leu Pro Ile Lys Arg Arg Glu Gln Ile Leu Ala Trp Ile Lys Glu1 5 10 15Glu Glu Ser Leu Arg Ile Ser Glu Ile Ser Lys Arg Leu

Asn Val Ser 20 25 30Glu Met Thr Val Tyr Arg Asp Ile Lys Pro Leu Val Glu Asn Gly Gln 35 40 45Val Ile Lys Thr Ala Gly Gly Ile Thr Leu Asn Lys Pro Lys Gln Gln 50 55 60Leu Gly Gln Leu Cys Val Val Cys Gly Lys Gly Ala Gly Ser Arg Leu65 70 75 80Ala Val Gln Ile Val Lys Asn Asp Ser Gln Ile Glu Gln Phe Cys Cys 85 90 95Val His Cys Ala Met Leu Arg Tyr Glu Lys Val Lys Asp Asp Val Ser 100 105 110Gln Ile Ile Cys Arg Asp Phe Leu Leu Asp Thr Thr Ile Ser Ala Lys 115 120 125Ala Ala Val Phe Leu Leu Asp Thr Asp Leu His Leu Asn Cys Cys Glu 130 135 140Pro His Ala Leu Pro Phe Ala Ser Glu Ala Glu Ala Arg Lys Phe Lys145 150 155 160Lys Gly Phe Gly Gly Gln Leu Leu Ser Phe Asp Asp Ala Ala Leu Leu 165 170 175Val Gln Lys Thr Met Lys Asn Ser Cys Cys Ser Leu Lys Thr 180 185 190332295DNABacillus licheniformis DSM 13gene(1)..(2295)nasB - assimilatory nitrate reductase (electron transfer subunit) - BLi00482 33atg ggg aaa caa agg ctt gtt tta ata gga aac ggc atg gca ggg gtt 48Met Gly Lys Gln Arg Leu Val Leu Ile Gly Asn Gly Met Ala Gly Val1 5 10 15cgc acg gcc gaa gag att tca cta cac ggc ggg agc cgg ttt gag atc 96Arg Thr Ala Glu Glu Ile Ser Leu His Gly Gly Ser Arg Phe Glu Ile 20 25 30gtc atc atc ggc agc gaa cca cac ccg ggc tat aac cgg att ctg ctg 144Val Ile Ile Gly Ser Glu Pro His Pro Gly Tyr Asn Arg Ile Leu Leu 35 40 45tct tct gtc ctt cag gga gaa acg gat tgg aac gat gtg atg aca aaa 192Ser Ser Val Leu Gln Gly Glu Thr Asp Trp Asn Asp Val Met Thr Lys 50 55 60agc cgg agc tgg tat gaa gaa aac ggg att acc ctt tat aca ggg gaa 240Ser Arg Ser Trp Tyr Glu Glu Asn Gly Ile Thr Leu Tyr Thr Gly Glu65 70 75 80acg gct gtc gca atc gat acg gtt aat caa acg gtg gcg aca gat caa 288Thr Ala Val Ala Ile Asp Thr Val Asn Gln Thr Val Ala Thr Asp Gln 85 90 95aac cgc gaa atc gct tat gac aaa ctg atc att gcg acg ggc tct tcc 336Asn Arg Glu Ile Ala Tyr Asp Lys Leu Ile Ile Ala Thr Gly Ser Ser 100 105 110ccg ttt atc ctc ccg gtt cat ggt gca gac aaa gaa ggg gta tat ggt 384Pro Phe Ile Leu Pro Val His Gly Ala Asp Lys Glu Gly Val Tyr Gly 115 120 125ttt aga aca ata gac gat tgc cgg gca ttc ata aac gca tca aag cga 432Phe Arg Thr Ile Asp Asp Cys Arg Ala Phe Ile Asn Ala Ser Lys Arg 130 135 140ttt aaa aaa gcg gcc gtc atc ggc ggg gga ata ttg ggc ctt gag gcg 480Phe Lys Lys Ala Ala Val Ile Gly Gly Gly Ile Leu Gly Leu Glu Ala145 150 155 160gcg cgg gga ttg gca aac ctc ggg atg aag gtt gat gtg gtt cac cac 528Ala Arg Gly Leu Ala Asn Leu Gly Met Lys Val Asp Val Val His His 165 170 175tct tcc tcc atc atg cag act cag ctt gat ccg ccg gct tct gcc atg 576Ser Ser Ser Ile Met Gln Thr Gln Leu Asp Pro Pro Ala Ser Ala Met 180 185 190ctc cag aaa gag ctg gag cgt caa ggg atc cat ttt ctt ttg gag aaa 624Leu Gln Lys Glu Leu Glu Arg Gln Gly Ile His Phe Leu Leu Glu Lys 195 200 205gat aca gag gcg att tta gga aca tcg cgc gcc gag ggt gtg cgg ttt 672Asp Thr Glu Ala Ile Leu Gly Thr Ser Arg Ala Glu Gly Val Arg Phe 210 215 220aaa gac ggg acg aag atc agc gcc gat ctc gtc gtc atg gct gcc ggt 720Lys Asp Gly Thr Lys Ile Ser Ala Asp Leu Val Val Met Ala Ala Gly225 230 235 240gtc aga ccg aat atc gaa ctg gcc aag gcg agc ggt att tca acc aac 768Val Arg Pro Asn Ile Glu Leu Ala Lys Ala Ser Gly Ile Ser Thr Asn 245 250 255agg gcg atc atc gtc agc gac tat atg gag aca aat gtt cca aat gtc 816Arg Ala Ile Ile Val Ser Asp Tyr Met Glu Thr Asn Val Pro Asn Val 260 265 270tac gcc gtc ggt gaa tgc gcg gag cat aac gga acg gtc tac ggc tta 864Tyr Ala Val Gly Glu Cys Ala Glu His Asn Gly Thr Val Tyr Gly Leu 275 280 285gtc aag cca ctg tat gag cag ggg aaa gtg ctg gcc aaa cat att tgc 912Val Lys Pro Leu Tyr Glu Gln Gly Lys Val Leu Ala Lys His Ile Cys 290 295 300ggc ctc gaa tgt tcc ggt tat cag ggc tcc gtt cag tcc gcg gct tta 960Gly Leu Glu Cys Ser Gly Tyr Gln Gly Ser Val Gln Ser Ala Ala Leu305 310 315 320aaa atg tcc gga atc gac gtc ttt tcg gca ggg aaa atc aca gag gac 1008Lys Met Ser Gly Ile Asp Val Phe Ser Ala Gly Lys Ile Thr Glu Asp 325 330 335gac tcg acg acg gct att aaa ctt tta gac gaa gcg gcc gga gtc tat 1056Asp Ser Thr Thr Ala Ile Lys Leu Leu Asp Glu Ala Ala Gly Val Tyr 340 345 350aaa aaa gcg gtc ttt caa ggg gat aaa atg gcc ggc gtc att tta tac 1104Lys Lys Ala Val Phe Gln Gly Asp Lys Met Ala Gly Val Ile Leu Tyr 355 360 365gga gac acc agc ggc agc caa agg ctg ttg gaa agc atc atc aaa caa 1152Gly Asp Thr Ser Gly Ser Gln Arg Leu Leu Glu Ser Ile Ile Lys Gln 370 375 380agg gat att acc gtt gtc aaa aaa gag ctg ttt caa tcg gag gaa gac 1200Arg Asp Ile Thr Val Val Lys Lys Glu Leu Phe Gln Ser Glu Glu Asp385 390 395 400agt tca gtc gct tct atg gcg gtt cgt gaa acg atc tgc caa tgc aac 1248Ser Ser Val Ala Ser Met Ala Val Arg Glu Thr Ile Cys Gln Cys Asn 405 410 415gcg gtt tca aaa ggc atg atc atg gaa gcg ata cag acg cac ggt tta 1296Ala Val Ser Lys Gly Met Ile Met Glu Ala Ile Gln Thr His Gly Leu 420 425 430aag acg gcc gag gag gtc aaa cgc tgt aca aaa gct tcc ggg tca tgc 1344Lys Thr Ala Glu Glu Val Lys Arg Cys Thr Lys Ala Ser Gly Ser Cys 435 440 445ggc ggc tgc agg ccg ttg gtc gag gaa ctg ctc ctc cat acc gcc gaa 1392Gly Gly Cys Arg Pro Leu Val Glu Glu Leu Leu Leu His Thr Ala Glu 450 455 460cat gat ggt cac ata tct gcg gcg gaa caa ccg atg tgc gcc tgc act 1440His Asp Gly His Ile Ser Ala Ala Glu Gln Pro Met Cys Ala Cys Thr465 470 475 480tca ttc acg gaa gat gaa gtc gtg aac gaa ata cag atg aga aat ctt 1488Ser Phe Thr Glu Asp Glu Val Val Asn Glu Ile Gln Met Arg Asn Leu 485 490 495tct tcc gtt cac gaa gtc atc tcg gcg ctc ggc tgg aaa aat agc agc 1536Ser Ser Val His Glu Val Ile Ser Ala Leu Gly Trp Lys Asn Ser Ser 500 505 510gga tgc cgg att tgc gtg ccc gcg ctt cac tac tat tta aaa atg atc 1584Gly Cys Arg Ile Cys Val Pro Ala Leu His Tyr Tyr Leu Lys Met Ile 515 520 525cgt ccg gac atc gcc tat gag gag cac gaa gag gag acg gat gcc atc 1632Arg Pro Asp Ile Ala Tyr Glu Glu His Glu Glu Glu Thr Asp Ala Ile 530 535 540atc ccg caa atg tac gga ggc cgg acg aat gcc gaa gag ctg aag cgc 1680Ile Pro Gln Met Tyr Gly Gly Arg Thr Asn Ala Glu Glu Leu Lys Arg545 550 555 560att gca gga gtg att gaa aaa tat caa att caa gag gtg tac atg aca 1728Ile Ala Gly Val Ile Glu Lys Tyr Gln Ile Gln Glu Val Tyr Met Thr 565 570 575cac cat cag aga tta aag ctt gcc ggc atc aag ccg gag cat ata gag 1776His His Gln Arg Leu Lys Leu Ala Gly Ile Lys Pro Glu His Ile Glu 580 585 590cgc gtg aaa gaa gaa ctc ggc atg ccg tat tgc ctg ccg caa cag cag 1824Arg Val Lys Glu Glu Leu Gly Met Pro Tyr Cys Leu Pro Gln Gln Gln 595 600 605cgg atc gcc gcg gtt aaa acg tgc cca tgc gga tca ccg cag att caa 1872Arg Ile Ala Ala Val Lys Thr Cys Pro Cys Gly Ser Pro Gln Ile Gln 610 615 620aca ttg gca gca gac ttg gaa aaa gca gca gaa tcc ctc ctc att ccg 1920Thr Leu Ala Ala Asp Leu Glu Lys Ala Ala Glu Ser Leu Leu Ile Pro625 630 635 640gcc aag gtc tcc att ggg gtg tcg ggc tgt ttg gac gat tgc gtg tac 1968Ala Lys Val Ser Ile Gly Val Ser Gly Cys Leu Asp Asp Cys Val Tyr 645 650 655gct tca gtt cat gac atc gga ctt ctc aaa gta aac gga gga tgg gag 2016Ala Ser Val His Asp Ile Gly Leu Leu Lys Val Asn Gly Gly Trp Glu 660 665 670att tat gcg ggc ggc caa gga gga cag cat gct tcc ccg gga gag ctg 2064Ile Tyr Ala Gly Gly Gln Gly Gly Gln His Ala Ser Pro Gly Glu Leu 675 680 685ctt tca gta gcc gac tcg gcg gag gaa gcg ggt gaa ctc atg aaa ggc 2112Leu Ser Val Ala Asp Ser Ala Glu Glu Ala Gly Glu Leu Met Lys Gly 690 695 700ctg ctt caa tat tac cgc gaa aca gca aac tat ctt gag aag atc ggg 2160Leu Leu Gln Tyr Tyr Arg Glu Thr Ala Asn Tyr Leu Glu Lys Ile Gly705 710 715 720cat tgg att gaa cgc gcc ggc atc att cat atc cgg gaa gtt tta ttt 2208His Trp Ile Glu Arg Ala Gly Ile Ile His Ile Arg Glu Val Leu Phe 725 730 735gac aac gga ctt cgc ggc cag ctg ttc gaa cgg ctt gaa aaa gaa aag 2256Asp Asn Gly Leu Arg Gly Gln Leu Phe Glu Arg Leu Glu Lys Glu Lys 740 745 750cgg ctg gct cag caa gaa atg ata aaa ggg ctg atg tga 2295Arg Leu Ala Gln Gln Glu Met Ile Lys Gly Leu Met 755 76034764PRTBacillus licheniformis DSM 13 34Met Gly Lys Gln Arg Leu Val Leu Ile Gly Asn Gly Met Ala Gly Val1 5 10 15Arg Thr Ala Glu Glu Ile Ser Leu His Gly Gly Ser Arg Phe Glu Ile 20 25 30Val Ile Ile Gly Ser Glu Pro His Pro Gly Tyr Asn Arg Ile Leu Leu 35 40 45Ser Ser Val Leu Gln Gly Glu Thr Asp Trp Asn Asp Val Met Thr Lys 50 55 60Ser Arg Ser Trp Tyr Glu Glu Asn Gly Ile Thr Leu Tyr Thr Gly Glu65 70 75 80Thr Ala Val Ala Ile Asp Thr Val Asn Gln Thr Val Ala Thr Asp Gln 85 90 95Asn Arg Glu Ile Ala Tyr Asp Lys Leu Ile Ile Ala Thr Gly Ser Ser 100 105 110Pro Phe Ile Leu Pro Val His Gly Ala Asp Lys Glu Gly Val Tyr Gly 115 120 125Phe Arg Thr Ile Asp Asp Cys Arg Ala Phe Ile Asn Ala Ser Lys Arg 130 135 140Phe Lys Lys Ala Ala Val Ile Gly Gly Gly Ile Leu Gly Leu Glu Ala145 150 155 160Ala Arg Gly Leu Ala Asn Leu Gly Met Lys Val Asp Val Val His His 165 170 175Ser Ser Ser Ile Met Gln Thr Gln Leu Asp Pro Pro Ala Ser Ala Met 180 185 190Leu Gln Lys Glu Leu Glu Arg Gln Gly Ile His Phe Leu Leu Glu Lys 195 200 205Asp Thr Glu Ala Ile Leu Gly Thr Ser Arg Ala Glu Gly Val Arg Phe 210 215 220Lys Asp Gly Thr Lys Ile Ser Ala Asp Leu Val Val Met Ala Ala Gly225 230 235 240Val Arg Pro Asn Ile Glu Leu Ala Lys Ala Ser Gly Ile Ser Thr Asn 245 250 255Arg Ala Ile Ile Val Ser Asp Tyr Met Glu Thr Asn Val Pro Asn Val 260 265 270Tyr Ala Val Gly Glu Cys Ala Glu His Asn Gly Thr Val Tyr Gly Leu 275 280 285Val Lys Pro Leu Tyr Glu Gln Gly Lys Val Leu Ala Lys His Ile Cys 290 295 300Gly Leu Glu Cys Ser Gly Tyr Gln Gly Ser Val Gln Ser Ala Ala Leu305 310 315 320Lys Met Ser Gly Ile Asp Val Phe Ser Ala Gly Lys Ile Thr Glu Asp 325 330 335Asp Ser Thr Thr Ala Ile Lys Leu Leu Asp Glu Ala Ala Gly Val Tyr 340 345 350Lys Lys Ala Val Phe Gln Gly Asp Lys Met Ala Gly Val Ile Leu Tyr 355 360 365Gly Asp Thr Ser Gly Ser Gln Arg Leu Leu Glu Ser Ile Ile Lys Gln 370 375 380Arg Asp Ile Thr Val Val Lys Lys Glu Leu Phe Gln Ser Glu Glu Asp385 390 395 400Ser Ser Val Ala Ser Met Ala Val Arg Glu Thr Ile Cys Gln Cys Asn 405 410 415Ala Val Ser Lys Gly Met Ile Met Glu Ala Ile Gln Thr His Gly Leu 420 425 430Lys Thr Ala Glu Glu Val Lys Arg Cys Thr Lys Ala Ser Gly Ser Cys 435 440 445Gly Gly Cys Arg Pro Leu Val Glu Glu Leu Leu Leu His Thr Ala Glu 450 455 460His Asp Gly His Ile Ser Ala Ala Glu Gln Pro Met Cys Ala Cys Thr465 470 475 480Ser Phe Thr Glu Asp Glu Val Val Asn Glu Ile Gln Met Arg Asn Leu 485 490 495Ser Ser Val His Glu Val Ile Ser Ala Leu Gly Trp Lys Asn Ser Ser 500 505 510Gly Cys Arg Ile Cys Val Pro Ala Leu His Tyr Tyr Leu Lys Met Ile 515 520 525Arg Pro Asp Ile Ala Tyr Glu Glu His Glu Glu Glu Thr Asp Ala Ile 530 535 540Ile Pro Gln Met Tyr Gly Gly Arg Thr Asn Ala Glu Glu Leu Lys Arg545 550 555 560Ile Ala Gly Val Ile Glu Lys Tyr Gln Ile Gln Glu Val Tyr Met Thr 565 570 575His His Gln Arg Leu Lys Leu Ala Gly Ile Lys Pro Glu His Ile Glu 580 585 590Arg Val Lys Glu Glu Leu Gly Met Pro Tyr Cys Leu Pro Gln Gln Gln 595 600 605Arg Ile Ala Ala Val Lys Thr Cys Pro Cys Gly Ser Pro Gln Ile Gln 610 615 620Thr Leu Ala Ala Asp Leu Glu Lys Ala Ala Glu Ser Leu Leu Ile Pro625 630 635 640Ala Lys Val Ser Ile Gly Val Ser Gly Cys Leu Asp Asp Cys Val Tyr 645 650 655Ala Ser Val His Asp Ile Gly Leu Leu Lys Val Asn Gly Gly Trp Glu 660 665 670Ile Tyr Ala Gly Gly Gln Gly Gly Gln His Ala Ser Pro Gly Glu Leu 675 680 685Leu Ser Val Ala Asp Ser Ala Glu Glu Ala Gly Glu Leu Met Lys Gly 690 695 700Leu Leu Gln Tyr Tyr Arg Glu Thr Ala Asn Tyr Leu Glu Lys Ile Gly705 710 715 720His Trp Ile Glu Arg Ala Gly Ile Ile His Ile Arg Glu Val Leu Phe 725 730 735Asp Asn Gly Leu Arg Gly Gln Leu Phe Glu Arg Leu Glu Lys Glu Lys 740 745 750Arg Leu Ala Gln Gln Glu Met Ile Lys Gly Leu Met 755 760352160DNABacillus licheniformis DSM 13gene(1)..(2160)nasC - assimilatory nitrate reductase (catalytic subunit) - BLi00483 35atg aca gaa cga ttg cta aag tac ttt cgc aca aag caa aaa gaa gtg 48Met Thr Glu Arg Leu Leu Lys Tyr Phe Arg Thr Lys Gln Lys Glu Val1 5 10 15caa tcc gaa aaa acg tat gac acc caa tgt cca ttt tgc agc atg cag 96Gln Ser Glu Lys Thr Tyr Asp Thr Gln Cys Pro Phe Cys Ser Met Gln 20 25 30tgc aaa atg cag ctc att gaa cag acg att gtg aca aga aaa aaa tac 144Cys Lys Met Gln Leu Ile Glu Gln Thr Ile Val Thr Arg Lys Lys Tyr 35 40 45acg gcg gtc gga aaa gac aat ccg acg aca aag ggg cgg ctg tgc atg 192Thr Ala Val Gly Lys Asp Asn Pro Thr Thr Lys Gly Arg Leu Cys Met 50 55 60aaa ggg atg aac gcc cat cag cac gcg ttt cat tca gag aga atc acc 240Lys Gly Met Asn Ala His Gln His Ala Phe His Ser Glu Arg Ile Thr65 70 75 80cat ccg ctg atc aaa aaa aac ggc gaa ttt gtc cgt gta tca tgg gaa 288His Pro Leu Ile Lys Lys Asn Gly Glu Phe Val Arg Val Ser Trp Glu 85 90 95gaa gcg ctt cag tac atc aaa gaa caa gta aca gcg atc cag cat gaa 336Glu Ala Leu Gln Tyr Ile Lys Glu Gln Val Thr Ala Ile Gln His Glu 100 105 110gac ggc cgc gat gct ttg agt gta tac ggc agc gcg tcc att aca aac 384Asp Gly Arg Asp Ala Leu Ser Val Tyr Gly Ser Ala Ser Ile Thr Asn 115 120 125gaa gaa gcg tac ctg ctc ggg aag ttc gcc cgc gtc gcc ttg aag acg 432Glu Glu Ala Tyr Leu Leu Gly Lys Phe Ala Arg Val Ala Leu Lys Thr 130 135 140aag cat att gac tac aac ggc agg ctt tgc atg tcg gcg gcc gca aca 480Lys His Ile Asp Tyr Asn Gly Arg Leu Cys Met Ser Ala Ala Ala Thr145 150 155 160gct gca aat caa acc ttc gga atg gac cgg gga ctg aca aat gcc ctt 528Ala Ala Asn Gln Thr Phe Gly

Met Asp Arg Gly Leu Thr Asn Ala Leu 165 170 175tcc gag atc ccc cat gca agg gtc atc atc ctc gca ggt acg aat atc 576Ser Glu Ile Pro His Ala Arg Val Ile Ile Leu Ala Gly Thr Asn Ile 180 185 190gcc gag tgc cag ccg acg atc atg ccg tat ttt gaa gaa gcg aag caa 624Ala Glu Cys Gln Pro Thr Ile Met Pro Tyr Phe Glu Glu Ala Lys Gln 195 200 205aac agg gcg tac atc atc gtc att gat ccc cgc gaa acg gca acg gcc 672Asn Arg Ala Tyr Ile Ile Val Ile Asp Pro Arg Glu Thr Ala Thr Ala 210 215 220aag atc gcc gac ctg cat ttg aaa atc aag ccg gga aca gac gcc gcg 720Lys Ile Ala Asp Leu His Leu Lys Ile Lys Pro Gly Thr Asp Ala Ala225 230 235 240ctc gca aac gga atg ctg aaa atc atc att gaa gaa cag ctg aca gac 768Leu Ala Asn Gly Met Leu Lys Ile Ile Ile Glu Glu Gln Leu Thr Asp 245 250 255gga gct ttt atc aga gat aga gct gaa ggg ttt gaa gag gtc aag cgc 816Gly Ala Phe Ile Arg Asp Arg Ala Glu Gly Phe Glu Glu Val Lys Arg 260 265 270cat gta gca tct tta tcg ctt gaa gac att tcg gcc aaa acc ggg ata 864His Val Ala Ser Leu Ser Leu Glu Asp Ile Ser Ala Lys Thr Gly Ile 275 280 285caa atc gag cag atc aaa aaa gcc gcg gta aaa ttc gcc cgc gag gag 912Gln Ile Glu Gln Ile Lys Lys Ala Ala Val Lys Phe Ala Arg Glu Glu 290 295 300acg gga atg ctg ttc acg gca aga ggg gtt gaa cag cag act gac gga 960Thr Gly Met Leu Phe Thr Ala Arg Gly Val Glu Gln Gln Thr Asp Gly305 310 315 320acg gct gct gta aga aat tta ctg aat att cta ata tca acg ggg aaa 1008Thr Ala Ala Val Arg Asn Leu Leu Asn Ile Leu Ile Ser Thr Gly Lys 325 330 335atc ggc aaa tat gct tgc ggc tac ggc gcc gtc acc ggg caa gga aac 1056Ile Gly Lys Tyr Ala Cys Gly Tyr Gly Ala Val Thr Gly Gln Gly Asn 340 345 350ggg cag ggg gcg cgg gag cac ggg cag aaa gca gac cag ctg cca ggt 1104Gly Gln Gly Ala Arg Glu His Gly Gln Lys Ala Asp Gln Leu Pro Gly 355 360 365tac cgt tcg att gaa aat gaa gcc gac cgc gcc tat att gcg gat gtt 1152Tyr Arg Ser Ile Glu Asn Glu Ala Asp Arg Ala Tyr Ile Ala Asp Val 370 375 380tgg ggc ata aaa gct gaa gat ctt ccg gga aaa ggc gta tcc gct tat 1200Trp Gly Ile Lys Ala Glu Asp Leu Pro Gly Lys Gly Val Ser Ala Tyr385 390 395 400gaa atg atg aag aaa gtt cat gaa ggc gaa att aca gga atg ttt tta 1248Glu Met Met Lys Lys Val His Glu Gly Glu Ile Thr Gly Met Phe Leu 405 410 415atg tgt tca aac cct gtt gta tcc agc ccg acc gcc ggc ttt gtc aaa 1296Met Cys Ser Asn Pro Val Val Ser Ser Pro Thr Ala Gly Phe Val Lys 420 425 430tcc gct ttg ggt aaa tta aag ttt ttt gcc gcc gtt gat ttg ttt ctt 1344Ser Ala Leu Gly Lys Leu Lys Phe Phe Ala Ala Val Asp Leu Phe Leu 435 440 445tct gaa acg gca ata ctc gcg gat gtc gtt ctc cca gca tca tcc tat 1392Ser Glu Thr Ala Ile Leu Ala Asp Val Val Leu Pro Ala Ser Ser Tyr 450 455 460ctc gag gac gaa gga acg atg acc aat ctg gaa ggc aga gtg aca tta 1440Leu Glu Asp Glu Gly Thr Met Thr Asn Leu Glu Gly Arg Val Thr Leu465 470 475 480agg gaa gcc tca aag ccg tgt ccc ggt gaa gcg agg cat gac tgg caa 1488Arg Glu Ala Ser Lys Pro Cys Pro Gly Glu Ala Arg His Asp Trp Gln 485 490 495att att tgc gat atc gct aaa gcg ctt gga aaa ggc cgc tat ttt tca 1536Ile Ile Cys Asp Ile Ala Lys Ala Leu Gly Lys Gly Arg Tyr Phe Ser 500 505 510tat cga tcg gca gag gat gtt ttt aat gaa ttg aga aaa gcg agc cgc 1584Tyr Arg Ser Ala Glu Asp Val Phe Asn Glu Leu Arg Lys Ala Ser Arg 515 520 525ggc gga att gcc gac tat tcg ggc atc acc tat gaa cgt ctg aga aag 1632Gly Gly Ile Ala Asp Tyr Ser Gly Ile Thr Tyr Glu Arg Leu Arg Lys 530 535 540gaa ggc ggc att tta tgg ccc tgt cca gaa gag ggg cac cct gga aca 1680Glu Gly Gly Ile Leu Trp Pro Cys Pro Glu Glu Gly His Pro Gly Thr545 550 555 560cag cgc ctg ttt gaa tca tcc ttc tcc cac ccg gac aaa aaa gca aga 1728Gln Arg Leu Phe Glu Ser Ser Phe Ser His Pro Asp Lys Lys Ala Arg 565 570 575atg tgc gtc gtc ccg aac gaa cca aag gtc aaa aaa gat gta ccg act 1776Met Cys Val Val Pro Asn Glu Pro Lys Val Lys Lys Asp Val Pro Thr 580 585 590gag gag tat ccc ctt att ctg acg acc gga agg gtg atg tct cac tat 1824Glu Glu Tyr Pro Leu Ile Leu Thr Thr Gly Arg Val Met Ser His Tyr 595 600 605tta acc ggg gta cag acg aga agg agc gct gcg ctc aag gcg aga cac 1872Leu Thr Gly Val Gln Thr Arg Arg Ser Ala Ala Leu Lys Ala Arg His 610 615 620ttt gaa tca ttt atg gaa atc cat ccg aaa acg gcg gaa aag tac aat 1920Phe Glu Ser Phe Met Glu Ile His Pro Lys Thr Ala Glu Lys Tyr Asn625 630 635 640att ggc gac cgg gtg ctg gtc agg gtg gaa tca aaa aga ggc agc atc 1968Ile Gly Asp Arg Val Leu Val Arg Val Glu Ser Lys Arg Gly Ser Ile 645 650 655atc gtc agg agc aaa tgg tcg gaa gcc atc agg ccg gat acc gtt ttt 2016Ile Val Arg Ser Lys Trp Ser Glu Ala Ile Arg Pro Asp Thr Val Phe 660 665 670gta ccg ata cat tgg gcg gat tcg caa aat gtg aac gat ttg ata gcg 2064Val Pro Ile His Trp Ala Asp Ser Gln Asn Val Asn Asp Leu Ile Ala 675 680 685gaa acg ctc gat ccg tcc tgc agc atg ccg gga ttt aag gtt tgc tcc 2112Glu Thr Leu Asp Pro Ser Cys Ser Met Pro Gly Phe Lys Val Cys Ser 690 695 700gtg aaa att cgt ccg tca gtt gac aat ttt gtg aat aaa ttg tca taa 2160Val Lys Ile Arg Pro Ser Val Asp Asn Phe Val Asn Lys Leu Ser705 710 71536719PRTBacillus licheniformis DSM 13 36Met Thr Glu Arg Leu Leu Lys Tyr Phe Arg Thr Lys Gln Lys Glu Val1 5 10 15Gln Ser Glu Lys Thr Tyr Asp Thr Gln Cys Pro Phe Cys Ser Met Gln 20 25 30Cys Lys Met Gln Leu Ile Glu Gln Thr Ile Val Thr Arg Lys Lys Tyr 35 40 45Thr Ala Val Gly Lys Asp Asn Pro Thr Thr Lys Gly Arg Leu Cys Met 50 55 60Lys Gly Met Asn Ala His Gln His Ala Phe His Ser Glu Arg Ile Thr65 70 75 80His Pro Leu Ile Lys Lys Asn Gly Glu Phe Val Arg Val Ser Trp Glu 85 90 95Glu Ala Leu Gln Tyr Ile Lys Glu Gln Val Thr Ala Ile Gln His Glu 100 105 110Asp Gly Arg Asp Ala Leu Ser Val Tyr Gly Ser Ala Ser Ile Thr Asn 115 120 125Glu Glu Ala Tyr Leu Leu Gly Lys Phe Ala Arg Val Ala Leu Lys Thr 130 135 140Lys His Ile Asp Tyr Asn Gly Arg Leu Cys Met Ser Ala Ala Ala Thr145 150 155 160Ala Ala Asn Gln Thr Phe Gly Met Asp Arg Gly Leu Thr Asn Ala Leu 165 170 175Ser Glu Ile Pro His Ala Arg Val Ile Ile Leu Ala Gly Thr Asn Ile 180 185 190Ala Glu Cys Gln Pro Thr Ile Met Pro Tyr Phe Glu Glu Ala Lys Gln 195 200 205Asn Arg Ala Tyr Ile Ile Val Ile Asp Pro Arg Glu Thr Ala Thr Ala 210 215 220Lys Ile Ala Asp Leu His Leu Lys Ile Lys Pro Gly Thr Asp Ala Ala225 230 235 240Leu Ala Asn Gly Met Leu Lys Ile Ile Ile Glu Glu Gln Leu Thr Asp 245 250 255Gly Ala Phe Ile Arg Asp Arg Ala Glu Gly Phe Glu Glu Val Lys Arg 260 265 270His Val Ala Ser Leu Ser Leu Glu Asp Ile Ser Ala Lys Thr Gly Ile 275 280 285Gln Ile Glu Gln Ile Lys Lys Ala Ala Val Lys Phe Ala Arg Glu Glu 290 295 300Thr Gly Met Leu Phe Thr Ala Arg Gly Val Glu Gln Gln Thr Asp Gly305 310 315 320Thr Ala Ala Val Arg Asn Leu Leu Asn Ile Leu Ile Ser Thr Gly Lys 325 330 335Ile Gly Lys Tyr Ala Cys Gly Tyr Gly Ala Val Thr Gly Gln Gly Asn 340 345 350Gly Gln Gly Ala Arg Glu His Gly Gln Lys Ala Asp Gln Leu Pro Gly 355 360 365Tyr Arg Ser Ile Glu Asn Glu Ala Asp Arg Ala Tyr Ile Ala Asp Val 370 375 380Trp Gly Ile Lys Ala Glu Asp Leu Pro Gly Lys Gly Val Ser Ala Tyr385 390 395 400Glu Met Met Lys Lys Val His Glu Gly Glu Ile Thr Gly Met Phe Leu 405 410 415Met Cys Ser Asn Pro Val Val Ser Ser Pro Thr Ala Gly Phe Val Lys 420 425 430Ser Ala Leu Gly Lys Leu Lys Phe Phe Ala Ala Val Asp Leu Phe Leu 435 440 445Ser Glu Thr Ala Ile Leu Ala Asp Val Val Leu Pro Ala Ser Ser Tyr 450 455 460Leu Glu Asp Glu Gly Thr Met Thr Asn Leu Glu Gly Arg Val Thr Leu465 470 475 480Arg Glu Ala Ser Lys Pro Cys Pro Gly Glu Ala Arg His Asp Trp Gln 485 490 495Ile Ile Cys Asp Ile Ala Lys Ala Leu Gly Lys Gly Arg Tyr Phe Ser 500 505 510Tyr Arg Ser Ala Glu Asp Val Phe Asn Glu Leu Arg Lys Ala Ser Arg 515 520 525Gly Gly Ile Ala Asp Tyr Ser Gly Ile Thr Tyr Glu Arg Leu Arg Lys 530 535 540Glu Gly Gly Ile Leu Trp Pro Cys Pro Glu Glu Gly His Pro Gly Thr545 550 555 560Gln Arg Leu Phe Glu Ser Ser Phe Ser His Pro Asp Lys Lys Ala Arg 565 570 575Met Cys Val Val Pro Asn Glu Pro Lys Val Lys Lys Asp Val Pro Thr 580 585 590Glu Glu Tyr Pro Leu Ile Leu Thr Thr Gly Arg Val Met Ser His Tyr 595 600 605Leu Thr Gly Val Gln Thr Arg Arg Ser Ala Ala Leu Lys Ala Arg His 610 615 620Phe Glu Ser Phe Met Glu Ile His Pro Lys Thr Ala Glu Lys Tyr Asn625 630 635 640Ile Gly Asp Arg Val Leu Val Arg Val Glu Ser Lys Arg Gly Ser Ile 645 650 655Ile Val Arg Ser Lys Trp Ser Glu Ala Ile Arg Pro Asp Thr Val Phe 660 665 670Val Pro Ile His Trp Ala Asp Ser Gln Asn Val Asn Asp Leu Ile Ala 675 680 685Glu Thr Leu Asp Pro Ser Cys Ser Met Pro Gly Phe Lys Val Cys Ser 690 695 700Val Lys Ile Arg Pro Ser Val Asp Asn Phe Val Asn Lys Leu Ser705 710 715372421DNABacillus licheniformis DSM 13gene(1)..(2421)nasD - assimilatory nitrite reductase (subunit) - BLi00484 37gtg aaa aaa aag cag ctt gtc ctt gta ggg aac ggt atg gcc ggg gtc 48Val Lys Lys Lys Gln Leu Val Leu Val Gly Asn Gly Met Ala Gly Val1 5 10 15agg gcc att gaa gaa att ctt aaa att tca aac gaa gag ttt cag atc 96Arg Ala Ile Glu Glu Ile Leu Lys Ile Ser Asn Glu Glu Phe Gln Ile 20 25 30acg att ttc ggc agc gaa ccg cac cca aac tat aac cgg att ctt ctt 144Thr Ile Phe Gly Ser Glu Pro His Pro Asn Tyr Asn Arg Ile Leu Leu 35 40 45tcc aaa gtg ctt caa gga gat acg gat gtt aag gac att aca ctg aat 192Ser Lys Val Leu Gln Gly Asp Thr Asp Val Lys Asp Ile Thr Leu Asn 50 55 60gat tgg gat tgg tat gaa gaa aac ggc att cag ctg tat aca ggt gaa 240Asp Trp Asp Trp Tyr Glu Glu Asn Gly Ile Gln Leu Tyr Thr Gly Glu65 70 75 80gaa gtc gtt aaa gtt gat ccg gag tcc aaa acc gtc atc acg gat acg 288Glu Val Val Lys Val Asp Pro Glu Ser Lys Thr Val Ile Thr Asp Thr 85 90 95gga aga gtc cag ccg tat gat gaa ctg att ttg gcg aca ggt tct ctc 336Gly Arg Val Gln Pro Tyr Asp Glu Leu Ile Leu Ala Thr Gly Ser Leu 100 105 110cct ttt att ctg cca ttg ccc gga gcg gat aaa gaa ggt gta aca gct 384Pro Phe Ile Leu Pro Leu Pro Gly Ala Asp Lys Glu Gly Val Thr Ala 115 120 125ttc cgt gat atc aag gat acg gat atc atg ctc gaa gct tcc aaa acg 432Phe Arg Asp Ile Lys Asp Thr Asp Ile Met Leu Glu Ala Ser Lys Thr 130 135 140tac aaa aaa gca gca gtt atc ggc ggc ggg ctg ctg gga ctg gaa gca 480Tyr Lys Lys Ala Ala Val Ile Gly Gly Gly Leu Leu Gly Leu Glu Ala145 150 155 160gcg cgc gga ctt ctc aac ctt ggc atg gat gtt acc gtt gtc cac ctc 528Ala Arg Gly Leu Leu Asn Leu Gly Met Asp Val Thr Val Val His Leu 165 170 175gcg cca tat ttg atg gag cgt cag ctt gat gca gcc gca gga cgg ctt 576Ala Pro Tyr Leu Met Glu Arg Gln Leu Asp Ala Ala Ala Gly Arg Leu 180 185 190ttg caa aaa gag ctg gaa aag cag ggc atg aaa ttt tta ctg gaa aag 624Leu Gln Lys Glu Leu Glu Lys Gln Gly Met Lys Phe Leu Leu Glu Lys 195 200 205cag acc gaa gaa att tac gga gaa aca cgt gtc gaa ggc tta aaa ttc 672Gln Thr Glu Glu Ile Tyr Gly Glu Thr Arg Val Glu Gly Leu Lys Phe 210 215 220aaa gac ggc tca acg ctt gag gcc gat ttg gtc gtc atg gcg gtc gga 720Lys Asp Gly Ser Thr Leu Glu Ala Asp Leu Val Val Met Ala Val Gly225 230 235 240atc aag ccg aac gtt cag ctc ggc aaa gac tgc ggc ctt ccg gta aac 768Ile Lys Pro Asn Val Gln Leu Gly Lys Asp Cys Gly Leu Pro Val Asn 245 250 255aga gga atc gtc gtc aat gat tac atg gag acc gaa gtt ccc aac att 816Arg Gly Ile Val Val Asn Asp Tyr Met Glu Thr Glu Val Pro Asn Ile 260 265 270tac gct gtc gga gaa tgc gcg gag cat agg ggc att gcc tac gga ctt 864Tyr Ala Val Gly Glu Cys Ala Glu His Arg Gly Ile Ala Tyr Gly Leu 275 280 285gtc gcg cct ctt tat gaa cag gca aaa gtg ctt gcc aaa aaa atc tgc 912Val Ala Pro Leu Tyr Glu Gln Ala Lys Val Leu Ala Lys Lys Ile Cys 290 295 300ggc gtt gaa acg gct ccg tat gaa gga tct gtt tta tct acc cag ctg 960Gly Val Glu Thr Ala Pro Tyr Glu Gly Ser Val Leu Ser Thr Gln Leu305 310 315 320aag gtt tcc ggt gtt gag gtc ttt tct gcc ggt gac ttt aat gag gat 1008Lys Val Ser Gly Val Glu Val Phe Ser Ala Gly Asp Phe Asn Glu Asp 325 330 335gac gga gaa gac aaa aaa gcg ctc aaa gtg ttt gac gaa cag gac gga 1056Asp Gly Glu Asp Lys Lys Ala Leu Lys Val Phe Asp Glu Gln Asp Gly 340 345 350atc tac aaa aaa atc gtt ctg cgc ggc aat cag gtg atc ggc gcc gta 1104Ile Tyr Lys Lys Ile Val Leu Arg Gly Asn Gln Val Ile Gly Ala Val 355 360 365ctg ttc gga gac agc agc gaa gga aac agg ctg ttt tcc atg att caa 1152Leu Phe Gly Asp Ser Ser Glu Gly Asn Arg Leu Phe Ser Met Ile Gln 370 375 380aaa gga gcg gat att tct gat act tca aaa att tca atc ttg cag cct 1200Lys Gly Ala Asp Ile Ser Asp Thr Ser Lys Ile Ser Ile Leu Gln Pro385 390 395 400ttg aat cag gaa gaa ggc atg agc ttg acc gct gcg atg agc gac gat 1248Leu Asn Gln Glu Glu Gly Met Ser Leu Thr Ala Ala Met Ser Asp Asp 405 410 415gag atc gtg tgc ggc tgc aac ggc gtt tcg aaa gga atg atc att cag 1296Glu Ile Val Cys Gly Cys Asn Gly Val Ser Lys Gly Met Ile Ile Gln 420 425 430gcg atc aaa gag cag gga tgc aca tcg acg gat gaa att aaa gca tgc 1344Ala Ile Lys Glu Gln Gly Cys Thr Ser Thr Asp Glu Ile Lys Ala Cys 435 440 445aca ggc gca tcg cgc tct tgc gga gga tgt aag ccg ctt gtc gca gag 1392Thr Gly Ala Ser Arg Ser Cys Gly Gly Cys Lys Pro Leu Val Ala Glu 450 455 460att ctc cag cat acg ctc ggc tca gaa ttt gat gct tca gca caa aaa 1440Ile Leu Gln His Thr Leu Gly Ser Glu Phe Asp Ala Ser Ala Gln Lys465 470 475 480gaa gcg att tgc gga tgt acg gac ctt tca aga gac gaa ctc gtg gcc 1488Glu Ala Ile Cys Gly Cys Thr Asp Leu Ser Arg Asp Glu Leu Val Ala 485 490 495gaa atc aga gaa aaa ggg ctg acc cat aca aaa gaa gtc atg aac gtg 1536Glu Ile Arg Glu Lys Gly Leu Thr His Thr Lys Glu Val Met Asn Val 500 505 510ctc ggc tgg aaa acg gcg gaa ggc tgt tcc aaa tgc cgc cct gcg ctt 1584Leu Gly Trp Lys Thr Ala Glu Gly Cys Ser Lys Cys Arg Pro Ala Leu 515 520 525aac tac tac ctc ggc atg atc aat ccg aag gaa tat gaa gat

gaa aga 1632Asn Tyr Tyr Leu Gly Met Ile Asn Pro Lys Glu Tyr Glu Asp Glu Arg 530 535 540gaa tcc cgc ttc gtc aat gaa cgg atg cac gcc aac att caa aaa gac 1680Glu Ser Arg Phe Val Asn Glu Arg Met His Ala Asn Ile Gln Lys Asp545 550 555 560ggc aca tat tca gtc gta ccg cgg atg tac ggc ggg gtc acc aat tcg 1728Gly Thr Tyr Ser Val Val Pro Arg Met Tyr Gly Gly Val Thr Asn Ser 565 570 575aat gac ttg cgg cgc atc gcc gat gtc gtc gac aaa tat gaa att cct 1776Asn Asp Leu Arg Arg Ile Ala Asp Val Val Asp Lys Tyr Glu Ile Pro 580 585 590cta gtg aaa atg acc ggc ggc cag cgg atc gat ctg atc ggc gtc aaa 1824Leu Val Lys Met Thr Gly Gly Gln Arg Ile Asp Leu Ile Gly Val Lys 595 600 605aaa gag gat ctt ccg aaa gta tgg gct gat ctt gac atg cct tct gga 1872Lys Glu Asp Leu Pro Lys Val Trp Ala Asp Leu Asp Met Pro Ser Gly 610 615 620ttt gct tac gga aaa acg ctc aga acc gta aaa acg tgc gta ggc gaa 1920Phe Ala Tyr Gly Lys Thr Leu Arg Thr Val Lys Thr Cys Val Gly Glu625 630 635 640cag ttc tgc cgc ttc gga acg caa gat tca atg gct ctc gga att gac 1968Gln Phe Cys Arg Phe Gly Thr Gln Asp Ser Met Ala Leu Gly Ile Asp 645 650 655ctt gag aaa aaa ttt gaa ggg ctc tac acg cct cac aaa att aaa atg 2016Leu Glu Lys Lys Phe Glu Gly Leu Tyr Thr Pro His Lys Ile Lys Met 660 665 670gcg gtt tcc gct tgt ccg aga aac tgt gct gaa tcc ggc att aaa gat 2064Ala Val Ser Ala Cys Pro Arg Asn Cys Ala Glu Ser Gly Ile Lys Asp 675 680 685atc ggc gtt gtc ggc atc gac ggc ggc tgg gaa att tac gtc ggc gga 2112Ile Gly Val Val Gly Ile Asp Gly Gly Trp Glu Ile Tyr Val Gly Gly 690 695 700aac gga ggc aca cat ctg cgt gca ggc gat ctg tta atg aaa gtc aaa 2160Asn Gly Gly Thr His Leu Arg Ala Gly Asp Leu Leu Met Lys Val Lys705 710 715 720acg gct gaa gaa gtc atc gaa atc aca ggc gca tat ctc caa tac tat 2208Thr Ala Glu Glu Val Ile Glu Ile Thr Gly Ala Tyr Leu Gln Tyr Tyr 725 730 735cgt gaa aca gcc aac tat tta gag cgg aca tct gca tgg atc gaa cgt 2256Arg Glu Thr Ala Asn Tyr Leu Glu Arg Thr Ser Ala Trp Ile Glu Arg 740 745 750gtc ggc ttg tcc cac gtc caa tcc gta ttg aat gat aaa gaa aaa cgc 2304Val Gly Leu Ser His Val Gln Ser Val Leu Asn Asp Lys Glu Lys Arg 755 760 765aaa gag ctg aat gac cgc atc aat caa gcg ctg tct gtc cat aaa gac 2352Lys Glu Leu Asn Asp Arg Ile Asn Gln Ala Leu Ser Val His Lys Asp 770 775 780ccg tgg aaa gac ttt ttg gaa gac gga aag aca aca aga cag cta ttc 2400Pro Trp Lys Asp Phe Leu Glu Asp Gly Lys Thr Thr Arg Gln Leu Phe785 790 795 800gaa aat gtt gtc act tcc taa 2421Glu Asn Val Val Thr Ser 80538806PRTBacillus licheniformis DSM 13misc_feature(1)..(1)Xaa can be Val or Met. 38Xaa Lys Lys Lys Gln Leu Val Leu Val Gly Asn Gly Met Ala Gly Val1 5 10 15Arg Ala Ile Glu Glu Ile Leu Lys Ile Ser Asn Glu Glu Phe Gln Ile 20 25 30Thr Ile Phe Gly Ser Glu Pro His Pro Asn Tyr Asn Arg Ile Leu Leu 35 40 45Ser Lys Val Leu Gln Gly Asp Thr Asp Val Lys Asp Ile Thr Leu Asn 50 55 60Asp Trp Asp Trp Tyr Glu Glu Asn Gly Ile Gln Leu Tyr Thr Gly Glu65 70 75 80Glu Val Val Lys Val Asp Pro Glu Ser Lys Thr Val Ile Thr Asp Thr 85 90 95Gly Arg Val Gln Pro Tyr Asp Glu Leu Ile Leu Ala Thr Gly Ser Leu 100 105 110Pro Phe Ile Leu Pro Leu Pro Gly Ala Asp Lys Glu Gly Val Thr Ala 115 120 125Phe Arg Asp Ile Lys Asp Thr Asp Ile Met Leu Glu Ala Ser Lys Thr 130 135 140Tyr Lys Lys Ala Ala Val Ile Gly Gly Gly Leu Leu Gly Leu Glu Ala145 150 155 160Ala Arg Gly Leu Leu Asn Leu Gly Met Asp Val Thr Val Val His Leu 165 170 175Ala Pro Tyr Leu Met Glu Arg Gln Leu Asp Ala Ala Ala Gly Arg Leu 180 185 190Leu Gln Lys Glu Leu Glu Lys Gln Gly Met Lys Phe Leu Leu Glu Lys 195 200 205Gln Thr Glu Glu Ile Tyr Gly Glu Thr Arg Val Glu Gly Leu Lys Phe 210 215 220Lys Asp Gly Ser Thr Leu Glu Ala Asp Leu Val Val Met Ala Val Gly225 230 235 240Ile Lys Pro Asn Val Gln Leu Gly Lys Asp Cys Gly Leu Pro Val Asn 245 250 255Arg Gly Ile Val Val Asn Asp Tyr Met Glu Thr Glu Val Pro Asn Ile 260 265 270Tyr Ala Val Gly Glu Cys Ala Glu His Arg Gly Ile Ala Tyr Gly Leu 275 280 285Val Ala Pro Leu Tyr Glu Gln Ala Lys Val Leu Ala Lys Lys Ile Cys 290 295 300Gly Val Glu Thr Ala Pro Tyr Glu Gly Ser Val Leu Ser Thr Gln Leu305 310 315 320Lys Val Ser Gly Val Glu Val Phe Ser Ala Gly Asp Phe Asn Glu Asp 325 330 335Asp Gly Glu Asp Lys Lys Ala Leu Lys Val Phe Asp Glu Gln Asp Gly 340 345 350Ile Tyr Lys Lys Ile Val Leu Arg Gly Asn Gln Val Ile Gly Ala Val 355 360 365Leu Phe Gly Asp Ser Ser Glu Gly Asn Arg Leu Phe Ser Met Ile Gln 370 375 380Lys Gly Ala Asp Ile Ser Asp Thr Ser Lys Ile Ser Ile Leu Gln Pro385 390 395 400Leu Asn Gln Glu Glu Gly Met Ser Leu Thr Ala Ala Met Ser Asp Asp 405 410 415Glu Ile Val Cys Gly Cys Asn Gly Val Ser Lys Gly Met Ile Ile Gln 420 425 430Ala Ile Lys Glu Gln Gly Cys Thr Ser Thr Asp Glu Ile Lys Ala Cys 435 440 445Thr Gly Ala Ser Arg Ser Cys Gly Gly Cys Lys Pro Leu Val Ala Glu 450 455 460Ile Leu Gln His Thr Leu Gly Ser Glu Phe Asp Ala Ser Ala Gln Lys465 470 475 480Glu Ala Ile Cys Gly Cys Thr Asp Leu Ser Arg Asp Glu Leu Val Ala 485 490 495Glu Ile Arg Glu Lys Gly Leu Thr His Thr Lys Glu Val Met Asn Val 500 505 510Leu Gly Trp Lys Thr Ala Glu Gly Cys Ser Lys Cys Arg Pro Ala Leu 515 520 525Asn Tyr Tyr Leu Gly Met Ile Asn Pro Lys Glu Tyr Glu Asp Glu Arg 530 535 540Glu Ser Arg Phe Val Asn Glu Arg Met His Ala Asn Ile Gln Lys Asp545 550 555 560Gly Thr Tyr Ser Val Val Pro Arg Met Tyr Gly Gly Val Thr Asn Ser 565 570 575Asn Asp Leu Arg Arg Ile Ala Asp Val Val Asp Lys Tyr Glu Ile Pro 580 585 590Leu Val Lys Met Thr Gly Gly Gln Arg Ile Asp Leu Ile Gly Val Lys 595 600 605Lys Glu Asp Leu Pro Lys Val Trp Ala Asp Leu Asp Met Pro Ser Gly 610 615 620Phe Ala Tyr Gly Lys Thr Leu Arg Thr Val Lys Thr Cys Val Gly Glu625 630 635 640Gln Phe Cys Arg Phe Gly Thr Gln Asp Ser Met Ala Leu Gly Ile Asp 645 650 655Leu Glu Lys Lys Phe Glu Gly Leu Tyr Thr Pro His Lys Ile Lys Met 660 665 670Ala Val Ser Ala Cys Pro Arg Asn Cys Ala Glu Ser Gly Ile Lys Asp 675 680 685Ile Gly Val Val Gly Ile Asp Gly Gly Trp Glu Ile Tyr Val Gly Gly 690 695 700Asn Gly Gly Thr His Leu Arg Ala Gly Asp Leu Leu Met Lys Val Lys705 710 715 720Thr Ala Glu Glu Val Ile Glu Ile Thr Gly Ala Tyr Leu Gln Tyr Tyr 725 730 735Arg Glu Thr Ala Asn Tyr Leu Glu Arg Thr Ser Ala Trp Ile Glu Arg 740 745 750Val Gly Leu Ser His Val Gln Ser Val Leu Asn Asp Lys Glu Lys Arg 755 760 765Lys Glu Leu Asn Asp Arg Ile Asn Gln Ala Leu Ser Val His Lys Asp 770 775 780Pro Trp Lys Asp Phe Leu Glu Asp Gly Lys Thr Thr Arg Gln Leu Phe785 790 795 800Glu Asn Val Val Thr Ser 805391461DNABacillus licheniformis DSM 13gene(1)..(1461)nasF - uroporphyrin-III C-methyltransferase - BLi00486 39atg ggg aat ggg aaa gtt tat ttt gtc ggg gcc gga ccg ggc gat tca 48Met Gly Asn Gly Lys Val Tyr Phe Val Gly Ala Gly Pro Gly Asp Ser1 5 10 15gga ttg atc aca tta aaa gga aga acg ctg atc gag gcg gcg gat gtc 96Gly Leu Ile Thr Leu Lys Gly Arg Thr Leu Ile Glu Ala Ala Asp Val 20 25 30ata ttg tat gac agg ctc gtc aat tcc cgg ctg ctt gag cat gca aaa 144Ile Leu Tyr Asp Arg Leu Val Asn Ser Arg Leu Leu Glu His Ala Lys 35 40 45gca tca tgc gaa ttt gtc tac tgc ggc aag ctc ccg gac cgc cat tat 192Ala Ser Cys Glu Phe Val Tyr Cys Gly Lys Leu Pro Asp Arg His Tyr 50 55 60atg agg caa agt gaa atc aat gca ctg ctc att gaa aag ggg ctg aaa 240Met Arg Gln Ser Glu Ile Asn Ala Leu Leu Ile Glu Lys Gly Leu Lys65 70 75 80ggc ctc aaa gtg gtc cgg ctg aaa gga gga gac ccg agc gtg ttt ggc 288Gly Leu Lys Val Val Arg Leu Lys Gly Gly Asp Pro Ser Val Phe Gly 85 90 95aga gtc ggg gaa gaa gcg gcc gcc gtc agc gaa cac ggc atc ccg tac 336Arg Val Gly Glu Glu Ala Ala Ala Val Ser Glu His Gly Ile Pro Tyr 100 105 110gag atg gtg cca ggc att aca tcg gga atc gcc gct cct ctt tat gcc 384Glu Met Val Pro Gly Ile Thr Ser Gly Ile Ala Ala Pro Leu Tyr Ala 115 120 125gga ctg cct gtg acc cac cgc gat ttc gct tca tcc ttt gcc gtt gtc 432Gly Leu Pro Val Thr His Arg Asp Phe Ala Ser Ser Phe Ala Val Val 130 135 140acc gcc cat gat aaa tcg ggc aaa ccg gag atg aat tgg gaa ggg ctg 480Thr Ala His Asp Lys Ser Gly Lys Pro Glu Met Asn Trp Glu Gly Leu145 150 155 160gcc aag ggc gtc caa acc ctc gtc ttt tac atg ggg atc aaa aac ctt 528Ala Lys Gly Val Gln Thr Leu Val Phe Tyr Met Gly Ile Lys Asn Leu 165 170 175ggt tat ata tgc gag cag ctg atg aaa cac ggc aag tcc ccc ctt gtt 576Gly Tyr Ile Cys Glu Gln Leu Met Lys His Gly Lys Ser Pro Leu Val 180 185 190ccc gca atg gcc atc caa tgg ggc aca tgg ggc cgg cag cgg agt gtt 624Pro Ala Met Ala Ile Gln Trp Gly Thr Trp Gly Arg Gln Arg Ser Val 195 200 205aaa ggg aca ttg cag aat atc agg gag aaa ata acc gag cat cag atc 672Lys Gly Thr Leu Gln Asn Ile Arg Glu Lys Ile Thr Glu His Gln Ile 210 215 220aaa aat ccc gcg att atc gtc atc ggc gac atc gta tca ttc agg cag 720Lys Asn Pro Ala Ile Ile Val Ile Gly Asp Ile Val Ser Phe Arg Gln225 230 235 240aac agc tgg ttt gaa gaa aag cct ttg atc ggg ctg aac atc atg gca 768Asn Ser Trp Phe Glu Glu Lys Pro Leu Ile Gly Leu Asn Ile Met Ala 245 250 255gtc aaa agc tgt gaa gaa gaa caa att cca gtg gag gcg ctt cgt gaa 816Val Lys Ser Cys Glu Glu Glu Gln Ile Pro Val Glu Ala Leu Arg Glu 260 265 270caa ggc gct gaa gtt gtg gaa tgg ccg aaa tgg aaa tcg gcc gaa acg 864Gln Gly Ala Glu Val Val Glu Trp Pro Lys Trp Lys Ser Ala Glu Thr 275 280 285gtg cct gac gat gca ctt tta ggt cga atc gct tgt ttt gac agc ctg 912Val Pro Asp Asp Ala Leu Leu Gly Arg Ile Ala Cys Phe Asp Ser Leu 290 295 300ctg ttc acc tca agt gcg tca ctg cgc aga ttt ttc agc cgg ctt gct 960Leu Phe Thr Ser Ser Ala Ser Leu Arg Arg Phe Phe Ser Arg Leu Ala305 310 315 320gct ttg aag atc gat gtc agg gaa atg cgc gga agg ctt tac gct tca 1008Ala Leu Lys Ile Asp Val Arg Glu Met Arg Gly Arg Leu Tyr Ala Ser 325 330 335gat cag cat gta aaa gca gct ctt gaa gaa cgg gga ttt gaa gcc ttt 1056Asp Gln His Val Lys Ala Ala Leu Glu Glu Arg Gly Phe Glu Ala Phe 340 345 350ctg caa acg gaa atg ccg gtc tcg ggc agc tgt ttg ctg atc ggc agc 1104Leu Gln Thr Glu Met Pro Val Ser Gly Ser Cys Leu Leu Ile Gly Ser 355 360 365cgg gaa gcc gtg caa aat tac ccg cac tcc ggg tgc gag gta ttg att 1152Arg Glu Ala Val Gln Asn Tyr Pro His Ser Gly Cys Glu Val Leu Ile 370 375 380aca cat gac agg gtg atc gac agc cgc ttt acc tcg atc atc aaa cgg 1200Thr His Asp Arg Val Ile Asp Ser Arg Phe Thr Ser Ile Ile Lys Arg385 390 395 400gaa att gat gag ttt cct att gat ata gtg ttg ttt tca aat cga aca 1248Glu Ile Asp Glu Phe Pro Ile Asp Ile Val Leu Phe Ser Asn Arg Thr 405 410 415tgc gtc gag ctg ttc gta aac gag gcg gag aca gtc ggc ctc cat ccg 1296Cys Val Glu Leu Phe Val Asn Glu Ala Glu Thr Val Gly Leu His Pro 420 425 430gaa aaa acg gct gcc cgt ctt cag gcg ttc tgc ctc act gaa gaa gcg 1344Glu Lys Thr Ala Ala Arg Leu Gln Ala Phe Cys Leu Thr Glu Glu Ala 435 440 445ggg gag aga gct gca gcc gta ggg ttt cag cgc gtt cat gca gtg aat 1392Gly Glu Arg Ala Ala Ala Val Gly Phe Gln Arg Val His Ala Val Asn 450 455 460gat cgc gat cgc ctg ccg gat gtc atc aaa gag atc aac cga aca gcc 1440Asp Arg Asp Arg Leu Pro Asp Val Ile Lys Glu Ile Asn Arg Thr Ala465 470 475 480gaa ttt gtc tgt cag agc tga 1461Glu Phe Val Cys Gln Ser 48540486PRTBacillus licheniformis DSM 13 40Met Gly Asn Gly Lys Val Tyr Phe Val Gly Ala Gly Pro Gly Asp Ser1 5 10 15Gly Leu Ile Thr Leu Lys Gly Arg Thr Leu Ile Glu Ala Ala Asp Val 20 25 30Ile Leu Tyr Asp Arg Leu Val Asn Ser Arg Leu Leu Glu His Ala Lys 35 40 45Ala Ser Cys Glu Phe Val Tyr Cys Gly Lys Leu Pro Asp Arg His Tyr 50 55 60Met Arg Gln Ser Glu Ile Asn Ala Leu Leu Ile Glu Lys Gly Leu Lys65 70 75 80Gly Leu Lys Val Val Arg Leu Lys Gly Gly Asp Pro Ser Val Phe Gly 85 90 95Arg Val Gly Glu Glu Ala Ala Ala Val Ser Glu His Gly Ile Pro Tyr 100 105 110Glu Met Val Pro Gly Ile Thr Ser Gly Ile Ala Ala Pro Leu Tyr Ala 115 120 125Gly Leu Pro Val Thr His Arg Asp Phe Ala Ser Ser Phe Ala Val Val 130 135 140Thr Ala His Asp Lys Ser Gly Lys Pro Glu Met Asn Trp Glu Gly Leu145 150 155 160Ala Lys Gly Val Gln Thr Leu Val Phe Tyr Met Gly Ile Lys Asn Leu 165 170 175Gly Tyr Ile Cys Glu Gln Leu Met Lys His Gly Lys Ser Pro Leu Val 180 185 190Pro Ala Met Ala Ile Gln Trp Gly Thr Trp Gly Arg Gln Arg Ser Val 195 200 205Lys Gly Thr Leu Gln Asn Ile Arg Glu Lys Ile Thr Glu His Gln Ile 210 215 220Lys Asn Pro Ala Ile Ile Val Ile Gly Asp Ile Val Ser Phe Arg Gln225 230 235 240Asn Ser Trp Phe Glu Glu Lys Pro Leu Ile Gly Leu Asn Ile Met Ala 245 250 255Val Lys Ser Cys Glu Glu Glu Gln Ile Pro Val Glu Ala Leu Arg Glu 260 265 270Gln Gly Ala Glu Val Val Glu Trp Pro Lys Trp Lys Ser Ala Glu Thr 275 280 285Val Pro Asp Asp Ala Leu Leu Gly Arg Ile Ala Cys Phe Asp Ser Leu 290 295 300Leu Phe Thr Ser Ser Ala Ser Leu Arg Arg Phe Phe Ser Arg Leu Ala305 310 315 320Ala Leu Lys Ile Asp Val Arg Glu Met Arg Gly Arg Leu Tyr Ala Ser 325 330 335Asp Gln His Val Lys Ala Ala Leu Glu Glu Arg Gly Phe Glu Ala Phe 340 345 350Leu Gln Thr Glu Met Pro Val Ser Gly Ser Cys Leu Leu Ile Gly Ser 355 360 365Arg Glu Ala Val Gln Asn Tyr Pro His Ser Gly Cys Glu Val Leu Ile 370 375 380Thr His Asp Arg Val Ile Asp Ser Arg Phe Thr Ser Ile Ile Lys Arg385 390 395 400Glu Ile Asp Glu Phe Pro Ile Asp Ile Val Leu Phe

Ser Asn Arg Thr 405 410 415Cys Val Glu Leu Phe Val Asn Glu Ala Glu Thr Val Gly Leu His Pro 420 425 430Glu Lys Thr Ala Ala Arg Leu Gln Ala Phe Cys Leu Thr Glu Glu Ala 435 440 445Gly Glu Arg Ala Ala Ala Val Gly Phe Gln Arg Val His Ala Val Asn 450 455 460Asp Arg Asp Arg Leu Pro Asp Val Ile Lys Glu Ile Asn Arg Thr Ala465 470 475 480Glu Phe Val Cys Gln Ser 485411191DNABacillus licheniformis DSM 13gene(1)..(1191)hypothetical protein - unknown function - BLi00236 41atg aaa agt cta aac aac att aaa aat aaa att gtc ata gtt gga aca 48Met Lys Ser Leu Asn Asn Ile Lys Asn Lys Ile Val Ile Val Gly Thr1 5 10 15gtt gat agc cta ggt gca gcc att gta tgg aca ggg tta cca ctt tat 96Val Asp Ser Leu Gly Ala Ala Ile Val Trp Thr Gly Leu Pro Leu Tyr 20 25 30gca tat tcg att acg gga tct ttt tta tat aca tca tca tta ttt ttt 144Ala Tyr Ser Ile Thr Gly Ser Phe Leu Tyr Thr Ser Ser Leu Phe Phe 35 40 45act tca gca ata gcc cgt ata tta gca acc ttt ttt gga ggc tat tta 192Thr Ser Ala Ile Ala Arg Ile Leu Ala Thr Phe Phe Gly Gly Tyr Leu 50 55 60gca gat gta tca tcg agt aaa aga ata atg cta aga tgt tta agt att 240Ala Asp Val Ser Ser Ser Lys Arg Ile Met Leu Arg Cys Leu Ser Ile65 70 75 80aat tgg ttt ttt tat tca cta tta ttt ttg gga tta tca ctc ggt tta 288Asn Trp Phe Phe Tyr Ser Leu Leu Phe Leu Gly Leu Ser Leu Gly Leu 85 90 95act tgg att ata ttc cct atc atg ttg att tca caa tgt ttc agt acc 336Thr Trp Ile Ile Phe Pro Ile Met Leu Ile Ser Gln Cys Phe Ser Thr 100 105 110ata tcg atc atg aat caa aac ctt tgg ttt aat tca ctt tgc tca aaa 384Ile Ser Ile Met Asn Gln Asn Leu Trp Phe Asn Ser Leu Cys Ser Lys 115 120 125gag acc ctg gct tcc gaa ata agt ggt aga aac tca tgg ttt ttg att 432Glu Thr Leu Ala Ser Glu Ile Ser Gly Arg Asn Ser Trp Phe Leu Ile 130 135 140agt aaa aca gtt ggt ttc tca tta gga cca ctc atg tat cat tgg ctt 480Ser Lys Thr Val Gly Phe Ser Leu Gly Pro Leu Met Tyr His Trp Leu145 150 155 160aat gca tac tca tta att atc aat cta tta act ata ttg att gca ata 528Asn Ala Tyr Ser Leu Ile Ile Asn Leu Leu Thr Ile Leu Ile Ala Ile 165 170 175aca tta ata tta aca ata aaa gat tta ggt tat gta aat aat aaa aag 576Thr Leu Ile Leu Thr Ile Lys Asp Leu Gly Tyr Val Asn Asn Lys Lys 180 185 190aaa aaa gag agc att ttt tct tcg ttc aaa gaa gtt tat aaa aat caa 624Lys Lys Glu Ser Ile Phe Ser Ser Phe Lys Glu Val Tyr Lys Asn Gln 195 200 205tta tta aaa agg cta aat ata ata gaa att tta aat ggt tta act ttt 672Leu Leu Lys Arg Leu Asn Ile Ile Glu Ile Leu Asn Gly Leu Thr Phe 210 215 220cct gtt tta aca agc tta tcg ata tat att ctt gaa aag aat tta aat 720Pro Val Leu Thr Ser Leu Ser Ile Tyr Ile Leu Glu Lys Asn Leu Asn225 230 235 240gct tct tct ctt ata att tct ttg ttt tgg ttg ata gga gga gta gga 768Ala Ser Ser Leu Ile Ile Ser Leu Phe Trp Leu Ile Gly Gly Val Gly 245 250 255gcg att acg gct aac ttg atg ctt tcg aga ctt aaa tta ttt aaa tta 816Ala Ile Thr Ala Asn Leu Met Leu Ser Arg Leu Lys Leu Phe Lys Leu 260 265 270cag tat att cat caa ttt ata ata agt ttt gtt ctc gta gtt ggt gga 864Gln Tyr Ile His Gln Phe Ile Ile Ser Phe Val Leu Val Val Gly Gly 275 280 285ttg ttg tta atg acg ata gct agc tat tct ttt gtc tat tta ata gga 912Leu Leu Leu Met Thr Ile Ala Ser Tyr Ser Phe Val Tyr Leu Ile Gly 290 295 300ttt tct tta ttt act tta ggt acc cct ata ata aat aat tta tta agg 960Phe Ser Leu Phe Thr Leu Gly Thr Pro Ile Ile Asn Asn Leu Leu Arg305 310 315 320tca aaa gtg ttt att ttc gcc cca agt cat atg aaa gga aaa gta aca 1008Ser Lys Val Phe Ile Phe Ala Pro Ser His Met Lys Gly Lys Val Thr 325 330 335tca gtt ata acg tct tca tca gat tta ggc act att ata gcc tta tca 1056Ser Val Ile Thr Ser Ser Ser Asp Leu Gly Thr Ile Ile Ala Leu Ser 340 345 350ttg tct tgg att gtt gtt gaa aaa ttc ggg gta tta att ttt atg tct 1104Leu Ser Trp Ile Val Val Glu Lys Phe Gly Val Leu Ile Phe Met Ser 355 360 365ttt gtt att tta aca agt gta agt cga ata tgc ctt ttt ttc tac act 1152Phe Val Ile Leu Thr Ser Val Ser Arg Ile Cys Leu Phe Phe Tyr Thr 370 375 380cta cat aaa gta gaa aat aaa gtt gat gtg gag gga taa 1191Leu His Lys Val Glu Asn Lys Val Asp Val Glu Gly385 390 39542396PRTBacillus licheniformis DSM 13 42Met Lys Ser Leu Asn Asn Ile Lys Asn Lys Ile Val Ile Val Gly Thr1 5 10 15Val Asp Ser Leu Gly Ala Ala Ile Val Trp Thr Gly Leu Pro Leu Tyr 20 25 30Ala Tyr Ser Ile Thr Gly Ser Phe Leu Tyr Thr Ser Ser Leu Phe Phe 35 40 45Thr Ser Ala Ile Ala Arg Ile Leu Ala Thr Phe Phe Gly Gly Tyr Leu 50 55 60Ala Asp Val Ser Ser Ser Lys Arg Ile Met Leu Arg Cys Leu Ser Ile65 70 75 80Asn Trp Phe Phe Tyr Ser Leu Leu Phe Leu Gly Leu Ser Leu Gly Leu 85 90 95Thr Trp Ile Ile Phe Pro Ile Met Leu Ile Ser Gln Cys Phe Ser Thr 100 105 110Ile Ser Ile Met Asn Gln Asn Leu Trp Phe Asn Ser Leu Cys Ser Lys 115 120 125Glu Thr Leu Ala Ser Glu Ile Ser Gly Arg Asn Ser Trp Phe Leu Ile 130 135 140Ser Lys Thr Val Gly Phe Ser Leu Gly Pro Leu Met Tyr His Trp Leu145 150 155 160Asn Ala Tyr Ser Leu Ile Ile Asn Leu Leu Thr Ile Leu Ile Ala Ile 165 170 175Thr Leu Ile Leu Thr Ile Lys Asp Leu Gly Tyr Val Asn Asn Lys Lys 180 185 190Lys Lys Glu Ser Ile Phe Ser Ser Phe Lys Glu Val Tyr Lys Asn Gln 195 200 205Leu Leu Lys Arg Leu Asn Ile Ile Glu Ile Leu Asn Gly Leu Thr Phe 210 215 220Pro Val Leu Thr Ser Leu Ser Ile Tyr Ile Leu Glu Lys Asn Leu Asn225 230 235 240Ala Ser Ser Leu Ile Ile Ser Leu Phe Trp Leu Ile Gly Gly Val Gly 245 250 255Ala Ile Thr Ala Asn Leu Met Leu Ser Arg Leu Lys Leu Phe Lys Leu 260 265 270Gln Tyr Ile His Gln Phe Ile Ile Ser Phe Val Leu Val Val Gly Gly 275 280 285Leu Leu Leu Met Thr Ile Ala Ser Tyr Ser Phe Val Tyr Leu Ile Gly 290 295 300Phe Ser Leu Phe Thr Leu Gly Thr Pro Ile Ile Asn Asn Leu Leu Arg305 310 315 320Ser Lys Val Phe Ile Phe Ala Pro Ser His Met Lys Gly Lys Val Thr 325 330 335Ser Val Ile Thr Ser Ser Ser Asp Leu Gly Thr Ile Ile Ala Leu Ser 340 345 350Leu Ser Trp Ile Val Val Glu Lys Phe Gly Val Leu Ile Phe Met Ser 355 360 365Phe Val Ile Leu Thr Ser Val Ser Arg Ile Cys Leu Phe Phe Tyr Thr 370 375 380Leu His Lys Val Glu Asn Lys Val Asp Val Glu Gly385 390 395431584DNABacillus licheniformis DSM 13gene(1)..(1584)pckA - phosphoenolpyruvate carboxykinase - BLi03197 43atg aat tct gta gat tta aca act gac tta caa acg cta tta acg ggc 48Met Asn Ser Val Asp Leu Thr Thr Asp Leu Gln Thr Leu Leu Thr Gly1 5 10 15gaa aac gtt cag atg aac tta tcc gtt ccc cat ttg gtt gaa aag atc 96Glu Asn Val Gln Met Asn Leu Ser Val Pro His Leu Val Glu Lys Ile 20 25 30ctt gag cgc aaa gaa gga gtc ttg aca tct tcc gga gcc gtg cgg gca 144Leu Glu Arg Lys Glu Gly Val Leu Thr Ser Ser Gly Ala Val Arg Ala 35 40 45aca acc ggt aca tac acc ggc cgc tcg cca aaa gac aaa ttc att gtg 192Thr Thr Gly Thr Tyr Thr Gly Arg Ser Pro Lys Asp Lys Phe Ile Val 50 55 60gaa gaa gcg agc aca aag gac aaa atc gat tgg ggc gca gta aac cag 240Glu Glu Ala Ser Thr Lys Asp Lys Ile Asp Trp Gly Ala Val Asn Gln65 70 75 80ccg att tca gaa gcc gcc ttt gac cgg ctg tac gca aaa gtg ctc ggc 288Pro Ile Ser Glu Ala Ala Phe Asp Arg Leu Tyr Ala Lys Val Leu Gly 85 90 95tat tta aag gaa cgc gat gaa ttg tat gta ttt gaa ggg ttt gca gga 336Tyr Leu Lys Glu Arg Asp Glu Leu Tyr Val Phe Glu Gly Phe Ala Gly 100 105 110gca gac gaa aaa tac cgg ctt ccg att act gtt gtc aac gag ttc gcc 384Ala Asp Glu Lys Tyr Arg Leu Pro Ile Thr Val Val Asn Glu Phe Ala 115 120 125tgg cac aac ctg ttt gca aga cag ttg ttc atc aga cct gaa gaa cat 432Trp His Asn Leu Phe Ala Arg Gln Leu Phe Ile Arg Pro Glu Glu His 130 135 140gat aaa aaa cct gaa gaa cag ccg ttt acg att ctt tca gct ccg cat 480Asp Lys Lys Pro Glu Glu Gln Pro Phe Thr Ile Leu Ser Ala Pro His145 150 155 160ttc aaa gcc gat ccg ctc gtt gac gga acc aga tct gaa acg ttt atc 528Phe Lys Ala Asp Pro Leu Val Asp Gly Thr Arg Ser Glu Thr Phe Ile 165 170 175atc gtg tcg ttt gag aag cgg acg atc tta atc gga ggc acc gaa tat 576Ile Val Ser Phe Glu Lys Arg Thr Ile Leu Ile Gly Gly Thr Glu Tyr 180 185 190gcc gga gag atg aaa aaa tcg att ttt tct att atg aat ttt ctt ctg 624Ala Gly Glu Met Lys Lys Ser Ile Phe Ser Ile Met Asn Phe Leu Leu 195 200 205cct gaa caa aac att cta ccg atg cac tgt tcg gca aac att ggc gag 672Pro Glu Gln Asn Ile Leu Pro Met His Cys Ser Ala Asn Ile Gly Glu 210 215 220gaa ggc gga act gct ctc ttc ttc gga ttg tca ggt aca ggc aaa acg 720Glu Gly Gly Thr Ala Leu Phe Phe Gly Leu Ser Gly Thr Gly Lys Thr225 230 235 240acg ctt tcc gcc gat ccg aag cgg aag ctg atc ggc gac gac gaa cac 768Thr Leu Ser Ala Asp Pro Lys Arg Lys Leu Ile Gly Asp Asp Glu His 245 250 255ggc tgg tcc agc aca ggc gtt ttc aac att gaa gga ggc tgc tac gcc 816Gly Trp Ser Ser Thr Gly Val Phe Asn Ile Glu Gly Gly Cys Tyr Ala 260 265 270aaa tgc att aac tta agc gaa gaa aaa gaa ccg cag att tac aat gcg 864Lys Cys Ile Asn Leu Ser Glu Glu Lys Glu Pro Gln Ile Tyr Asn Ala 275 280 285atc cgc ttc ggt tcc gtt ctc gaa aat gtc gtg gtt gac agc aaa acc 912Ile Arg Phe Gly Ser Val Leu Glu Asn Val Val Val Asp Ser Lys Thr 290 295 300gga gag ccg gac tat tcc gac tca ttc tac acg gaa aac act aga gcg 960Gly Glu Pro Asp Tyr Ser Asp Ser Phe Tyr Thr Glu Asn Thr Arg Ala305 310 315 320gct tac ccg att gac gcg att gac aac atc gtc aag ccg agc atc gca 1008Ala Tyr Pro Ile Asp Ala Ile Asp Asn Ile Val Lys Pro Ser Ile Ala 325 330 335gcg cac ccg cag acc atc gta ttt ttg acc gca gat gct ttc ggc gtg 1056Ala His Pro Gln Thr Ile Val Phe Leu Thr Ala Asp Ala Phe Gly Val 340 345 350ctt cct ccg atc agc aag tta acg aaa gaa caa gcc atg tat cat ttc 1104Leu Pro Pro Ile Ser Lys Leu Thr Lys Glu Gln Ala Met Tyr His Phe 355 360 365ctg agc gga tat aca agc aag ctc gcc ggc acc gaa cgc ggg atc acc 1152Leu Ser Gly Tyr Thr Ser Lys Leu Ala Gly Thr Glu Arg Gly Ile Thr 370 375 380act cct gaa gcc aca ttc tcg acc tgc ttc ggt tca ccg ttc ctc ccg 1200Thr Pro Glu Ala Thr Phe Ser Thr Cys Phe Gly Ser Pro Phe Leu Pro385 390 395 400ctt ccg gca cat gtg tac gct gaa atg ctc gga aga aaa atc gat gag 1248Leu Pro Ala His Val Tyr Ala Glu Met Leu Gly Arg Lys Ile Asp Glu 405 410 415cac ggg gtc ggc gtc ttt ctc gtc aat aca ggc tgg acc ggc ggc gga 1296His Gly Val Gly Val Phe Leu Val Asn Thr Gly Trp Thr Gly Gly Gly 420 425 430tac gga acc ggg gaa aga atg agc ctc gca tat acg aga gcg atg gtt 1344Tyr Gly Thr Gly Glu Arg Met Ser Leu Ala Tyr Thr Arg Ala Met Val 435 440 445cag gcc gca ata gaa gga gaa ctg gat aaa gcc gaa atg agg act gac 1392Gln Ala Ala Ile Glu Gly Glu Leu Asp Lys Ala Glu Met Arg Thr Asp 450 455 460cgc att ttc gga ctt cat tca cct gtc cac gtt ccg ggt gtg cct gat 1440Arg Ile Phe Gly Leu His Ser Pro Val His Val Pro Gly Val Pro Asp465 470 475 480caa gtg ctt gac ccg gcc aaa act tgg aca gat gaa aac gaa tac gag 1488Gln Val Leu Asp Pro Ala Lys Thr Trp Thr Asp Glu Asn Glu Tyr Glu 485 490 495gaa aaa gcg atc cac ctg gcg aaa gca ttc aaa cag aat ttc aaa aag 1536Glu Lys Ala Ile His Leu Ala Lys Ala Phe Lys Gln Asn Phe Lys Lys 500 505 510ttc tcc aat acg gaa aac atc gaa aaa gcc gga gga ccg ctc gtt taa 1584Phe Ser Asn Thr Glu Asn Ile Glu Lys Ala Gly Gly Pro Leu Val 515 520 52544527PRTBacillus licheniformis DSM 13 44Met Asn Ser Val Asp Leu Thr Thr Asp Leu Gln Thr Leu Leu Thr Gly1 5 10 15Glu Asn Val Gln Met Asn Leu Ser Val Pro His Leu Val Glu Lys Ile 20 25 30Leu Glu Arg Lys Glu Gly Val Leu Thr Ser Ser Gly Ala Val Arg Ala 35 40 45Thr Thr Gly Thr Tyr Thr Gly Arg Ser Pro Lys Asp Lys Phe Ile Val 50 55 60Glu Glu Ala Ser Thr Lys Asp Lys Ile Asp Trp Gly Ala Val Asn Gln65 70 75 80Pro Ile Ser Glu Ala Ala Phe Asp Arg Leu Tyr Ala Lys Val Leu Gly 85 90 95Tyr Leu Lys Glu Arg Asp Glu Leu Tyr Val Phe Glu Gly Phe Ala Gly 100 105 110Ala Asp Glu Lys Tyr Arg Leu Pro Ile Thr Val Val Asn Glu Phe Ala 115 120 125Trp His Asn Leu Phe Ala Arg Gln Leu Phe Ile Arg Pro Glu Glu His 130 135 140Asp Lys Lys Pro Glu Glu Gln Pro Phe Thr Ile Leu Ser Ala Pro His145 150 155 160Phe Lys Ala Asp Pro Leu Val Asp Gly Thr Arg Ser Glu Thr Phe Ile 165 170 175Ile Val Ser Phe Glu Lys Arg Thr Ile Leu Ile Gly Gly Thr Glu Tyr 180 185 190Ala Gly Glu Met Lys Lys Ser Ile Phe Ser Ile Met Asn Phe Leu Leu 195 200 205Pro Glu Gln Asn Ile Leu Pro Met His Cys Ser Ala Asn Ile Gly Glu 210 215 220Glu Gly Gly Thr Ala Leu Phe Phe Gly Leu Ser Gly Thr Gly Lys Thr225 230 235 240Thr Leu Ser Ala Asp Pro Lys Arg Lys Leu Ile Gly Asp Asp Glu His 245 250 255Gly Trp Ser Ser Thr Gly Val Phe Asn Ile Glu Gly Gly Cys Tyr Ala 260 265 270Lys Cys Ile Asn Leu Ser Glu Glu Lys Glu Pro Gln Ile Tyr Asn Ala 275 280 285Ile Arg Phe Gly Ser Val Leu Glu Asn Val Val Val Asp Ser Lys Thr 290 295 300Gly Glu Pro Asp Tyr Ser Asp Ser Phe Tyr Thr Glu Asn Thr Arg Ala305 310 315 320Ala Tyr Pro Ile Asp Ala Ile Asp Asn Ile Val Lys Pro Ser Ile Ala 325 330 335Ala His Pro Gln Thr Ile Val Phe Leu Thr Ala Asp Ala Phe Gly Val 340 345 350Leu Pro Pro Ile Ser Lys Leu Thr Lys Glu Gln Ala Met Tyr His Phe 355 360 365Leu Ser Gly Tyr Thr Ser Lys Leu Ala Gly Thr Glu Arg Gly Ile Thr 370 375 380Thr Pro Glu Ala Thr Phe Ser Thr Cys Phe Gly Ser Pro Phe Leu Pro385 390 395 400Leu Pro Ala His Val Tyr Ala Glu Met Leu Gly Arg Lys Ile Asp Glu 405 410 415His Gly Val Gly Val Phe Leu Val Asn Thr Gly Trp Thr Gly Gly Gly 420 425 430Tyr Gly Thr Gly Glu Arg Met Ser Leu Ala Tyr Thr Arg Ala Met Val 435 440 445Gln Ala Ala Ile Glu Gly Glu Leu Asp Lys Ala Glu Met Arg Thr Asp 450 455 460Arg Ile Phe Gly Leu His Ser Pro Val His Val Pro Gly Val Pro

Asp465 470 475 480Gln Val Leu Asp Pro Ala Lys Thr Trp Thr Asp Glu Asn Glu Tyr Glu 485 490 495Glu Lys Ala Ile His Leu Ala Lys Ala Phe Lys Gln Asn Phe Lys Lys 500 505 510Phe Ser Asn Thr Glu Asn Ile Glu Lys Ala Gly Gly Pro Leu Val 515 520 525451590DNABacillus licheniformis DSM 13gene(1)..(1590)putative protein - putative malate synthase (EC 4.1.3.2) - BLi04208 45atg aaa aca gag gag aag aga att cat ata acc gga tcg ata aat ccg 48Met Lys Thr Glu Glu Lys Arg Ile His Ile Thr Gly Ser Ile Asn Pro1 5 10 15cat tat gaa gag att ttg aca aaa gaa gcg atg gag ttt atc gga cgt 96His Tyr Glu Glu Ile Leu Thr Lys Glu Ala Met Glu Phe Ile Gly Arg 20 25 30ctt gaa cgc cat ttc ggg gga aga aga cgg gaa ttg ctg cat ttg cgg 144Leu Glu Arg His Phe Gly Gly Arg Arg Arg Glu Leu Leu His Leu Arg 35 40 45acg atc agg cag gct gaa att gat caa ggg aag aac cct tgt ttt ttg 192Thr Ile Arg Gln Ala Glu Ile Asp Gln Gly Lys Asn Pro Cys Phe Leu 50 55 60gcg gaa acg aag cac atc cgc gag gcg gac tgg aca gtc gcg ccg ctc 240Ala Glu Thr Lys His Ile Arg Glu Ala Asp Trp Thr Val Ala Pro Leu65 70 75 80ccc cgg gat ttg gag gac cgc agg gtt gaa atc act ggc ccg gtc gac 288Pro Arg Asp Leu Glu Asp Arg Arg Val Glu Ile Thr Gly Pro Val Asp 85 90 95aga aaa atg gtc atc aac gct tta aat tca gga gcg aaa gtc ttt atg 336Arg Lys Met Val Ile Asn Ala Leu Asn Ser Gly Ala Lys Val Phe Met 100 105 110gcc gac ttt gaa gat gcg aac tcg ccg acg tgg aaa aac tgc att gaa 384Ala Asp Phe Glu Asp Ala Asn Ser Pro Thr Trp Lys Asn Cys Ile Glu 115 120 125ggc cag gtt aat ttg cgg gat gcg gtc cgc ggg acg att tct ttt acg 432Gly Gln Val Asn Leu Arg Asp Ala Val Arg Gly Thr Ile Ser Phe Thr 130 135 140aat gaa cag ggc aaa cgc tat gag ctg aaa gag aac ccc gcc gtt atc 480Asn Glu Gln Gly Lys Arg Tyr Glu Leu Lys Glu Asn Pro Ala Val Ile145 150 155 160atg gtc agg ccg agg gga tgg cac ttg gaa gaa aag cat gtg ctg atc 528Met Val Arg Pro Arg Gly Trp His Leu Glu Glu Lys His Val Leu Ile 165 170 175ggc ggc agg ccc gtg tca gca agc ctg ttc gat ttt ggt ctt tat ttt 576Gly Gly Arg Pro Val Ser Ala Ser Leu Phe Asp Phe Gly Leu Tyr Phe 180 185 190ttc cat aac gca aaa gag ctg atc aaa aaa gga agc ggc cct tat ttc 624Phe His Asn Ala Lys Glu Leu Ile Lys Lys Gly Ser Gly Pro Tyr Phe 195 200 205tac ctt cca aaa atc gaa aac cac ctg gaa gca agg ctg tgg aac gac 672Tyr Leu Pro Lys Ile Glu Asn His Leu Glu Ala Arg Leu Trp Asn Asp 210 215 220gta ttt gtt tat gcg caa aac gac ttg cgc att ccg cag ggc acg att 720Val Phe Val Tyr Ala Gln Asn Asp Leu Arg Ile Pro Gln Gly Thr Ile225 230 235 240aaa gcg acg gtt ctc gtt gaa acg atc ctg gcg gcc ttt gaa atg gat 768Lys Ala Thr Val Leu Val Glu Thr Ile Leu Ala Ala Phe Glu Met Asp 245 250 255gaa atc ctc tac gaa ttg aag gaa cat tca gcc ggg ctt aac tgc ggc 816Glu Ile Leu Tyr Glu Leu Lys Glu His Ser Ala Gly Leu Asn Cys Gly 260 265 270agg tgg gat tat att ttc agc ttt att aag aaa ttc agg aac agg gat 864Arg Trp Asp Tyr Ile Phe Ser Phe Ile Lys Lys Phe Arg Asn Arg Asp 275 280 285gaa gtc att ctt cca gat cgc tca gcc gtg acg atg aca gtt ccc aac 912Glu Val Ile Leu Pro Asp Arg Ser Ala Val Thr Met Thr Val Pro Asn 290 295 300atg aga gcc tac tcc ctc ttg gcg att caa acg tgt cat aaa cgc aac 960Met Arg Ala Tyr Ser Leu Leu Ala Ile Gln Thr Cys His Lys Arg Asn305 310 315 320gtc cat gcg att ggg gga atg gcg gcg caa att ccc gct aaa gac aaa 1008Val His Ala Ile Gly Gly Met Ala Ala Gln Ile Pro Ala Lys Asp Lys 325 330 335cct gag atc aat gaa gcc gca ttt gcc aaa atc cgc gaa gat aag gaa 1056Pro Glu Ile Asn Glu Ala Ala Phe Ala Lys Ile Arg Glu Asp Lys Glu 340 345 350cgt gaa gta aaa gac ggc cat gac gga aca tgg atc gcg cat ccc ggc 1104Arg Glu Val Lys Asp Gly His Asp Gly Thr Trp Ile Ala His Pro Gly 355 360 365atg att caa acg gcg ctc gat gta ttc aat cag cat atg ccg tcg gcg 1152Met Ile Gln Thr Ala Leu Asp Val Phe Asn Gln His Met Pro Ser Ala 370 375 380aac cag att cat aga aag cgg aaa gac gta aag gtc gag gaa aaa gac 1200Asn Gln Ile His Arg Lys Arg Lys Asp Val Lys Val Glu Glu Lys Asp385 390 395 400ttg ctg gaa gta ccg aaa ggc aga att aca gaa gca ggc gtc cgt acc 1248Leu Leu Glu Val Pro Lys Gly Arg Ile Thr Glu Ala Gly Val Arg Thr 405 410 415aat atc agc gcc ggc atc cgc tat tta gct tca tgg ctg tcg ggc aga 1296Asn Ile Ser Ala Gly Ile Arg Tyr Leu Ala Ser Trp Leu Ser Gly Arg 420 425 430ggc gcc gct ccg atc gac cac tta atg gag gac gct gca acg gcg gag 1344Gly Ala Ala Pro Ile Asp His Leu Met Glu Asp Ala Ala Thr Ala Glu 435 440 445att tcc aga gcc cag ctg tgg caa tgg att cgc cat ccg aaa gga gtt 1392Ile Ser Arg Ala Gln Leu Trp Gln Trp Ile Arg His Pro Lys Gly Val 450 455 460ctt gac gat ggc cgg aaa gtc acc gcg gaa atg gtc aag atg atg aaa 1440Leu Asp Asp Gly Arg Lys Val Thr Ala Glu Met Val Lys Met Met Lys465 470 475 480gag aaa gaa atg cag aaa atc atc gat gaa gca ggc gtg gaa cat ttt 1488Glu Lys Glu Met Gln Lys Ile Ile Asp Glu Ala Gly Val Glu His Phe 485 490 495cgg agc ggc cgg ttt gaa gag gcc gga cag tta ttt gaa cag ctg att 1536Arg Ser Gly Arg Phe Glu Glu Ala Gly Gln Leu Phe Glu Gln Leu Ile 500 505 510ttg gat gat gaa ttc gcg gag ttt cta acg ata ccg gct tat gaa aat 1584Leu Asp Asp Glu Phe Ala Glu Phe Leu Thr Ile Pro Ala Tyr Glu Asn 515 520 525ata taa 1590Ile 46529PRTBacillus licheniformis DSM 13 46Met Lys Thr Glu Glu Lys Arg Ile His Ile Thr Gly Ser Ile Asn Pro1 5 10 15His Tyr Glu Glu Ile Leu Thr Lys Glu Ala Met Glu Phe Ile Gly Arg 20 25 30Leu Glu Arg His Phe Gly Gly Arg Arg Arg Glu Leu Leu His Leu Arg 35 40 45Thr Ile Arg Gln Ala Glu Ile Asp Gln Gly Lys Asn Pro Cys Phe Leu 50 55 60Ala Glu Thr Lys His Ile Arg Glu Ala Asp Trp Thr Val Ala Pro Leu65 70 75 80Pro Arg Asp Leu Glu Asp Arg Arg Val Glu Ile Thr Gly Pro Val Asp 85 90 95Arg Lys Met Val Ile Asn Ala Leu Asn Ser Gly Ala Lys Val Phe Met 100 105 110Ala Asp Phe Glu Asp Ala Asn Ser Pro Thr Trp Lys Asn Cys Ile Glu 115 120 125Gly Gln Val Asn Leu Arg Asp Ala Val Arg Gly Thr Ile Ser Phe Thr 130 135 140Asn Glu Gln Gly Lys Arg Tyr Glu Leu Lys Glu Asn Pro Ala Val Ile145 150 155 160Met Val Arg Pro Arg Gly Trp His Leu Glu Glu Lys His Val Leu Ile 165 170 175Gly Gly Arg Pro Val Ser Ala Ser Leu Phe Asp Phe Gly Leu Tyr Phe 180 185 190Phe His Asn Ala Lys Glu Leu Ile Lys Lys Gly Ser Gly Pro Tyr Phe 195 200 205Tyr Leu Pro Lys Ile Glu Asn His Leu Glu Ala Arg Leu Trp Asn Asp 210 215 220Val Phe Val Tyr Ala Gln Asn Asp Leu Arg Ile Pro Gln Gly Thr Ile225 230 235 240Lys Ala Thr Val Leu Val Glu Thr Ile Leu Ala Ala Phe Glu Met Asp 245 250 255Glu Ile Leu Tyr Glu Leu Lys Glu His Ser Ala Gly Leu Asn Cys Gly 260 265 270Arg Trp Asp Tyr Ile Phe Ser Phe Ile Lys Lys Phe Arg Asn Arg Asp 275 280 285Glu Val Ile Leu Pro Asp Arg Ser Ala Val Thr Met Thr Val Pro Asn 290 295 300Met Arg Ala Tyr Ser Leu Leu Ala Ile Gln Thr Cys His Lys Arg Asn305 310 315 320Val His Ala Ile Gly Gly Met Ala Ala Gln Ile Pro Ala Lys Asp Lys 325 330 335Pro Glu Ile Asn Glu Ala Ala Phe Ala Lys Ile Arg Glu Asp Lys Glu 340 345 350Arg Glu Val Lys Asp Gly His Asp Gly Thr Trp Ile Ala His Pro Gly 355 360 365Met Ile Gln Thr Ala Leu Asp Val Phe Asn Gln His Met Pro Ser Ala 370 375 380Asn Gln Ile His Arg Lys Arg Lys Asp Val Lys Val Glu Glu Lys Asp385 390 395 400Leu Leu Glu Val Pro Lys Gly Arg Ile Thr Glu Ala Gly Val Arg Thr 405 410 415Asn Ile Ser Ala Gly Ile Arg Tyr Leu Ala Ser Trp Leu Ser Gly Arg 420 425 430Gly Ala Ala Pro Ile Asp His Leu Met Glu Asp Ala Ala Thr Ala Glu 435 440 445Ile Ser Arg Ala Gln Leu Trp Gln Trp Ile Arg His Pro Lys Gly Val 450 455 460Leu Asp Asp Gly Arg Lys Val Thr Ala Glu Met Val Lys Met Met Lys465 470 475 480Glu Lys Glu Met Gln Lys Ile Ile Asp Glu Ala Gly Val Glu His Phe 485 490 495Arg Ser Gly Arg Phe Glu Glu Ala Gly Gln Leu Phe Glu Gln Leu Ile 500 505 510Leu Asp Asp Glu Phe Ala Glu Phe Leu Thr Ile Pro Ala Tyr Glu Asn 515 520 525Ile 471350DNABacillus licheniformis DSM 13gene(1)..(1350)putative protein - putative hydrolase - BLi02233 47atg caa aaa acg ctt ttt caa aat gct acg att gtg acg ctt gat ccg 48Met Gln Lys Thr Leu Phe Gln Asn Ala Thr Ile Val Thr Leu Asp Pro1 5 10 15cag ctt cat acg ctc gaa aaa ggg gac atg ctg att gaa ggc tcc gtc 96Gln Leu His Thr Leu Glu Lys Gly Asp Met Leu Ile Glu Gly Ser Val 20 25 30att ctg gac gtt gcc gag tcg att gaa gcc gga gac gct gag att gtg 144Ile Leu Asp Val Ala Glu Ser Ile Glu Ala Gly Asp Ala Glu Ile Val 35 40 45gat gcc tcc aat atg ctg atc atg cct gga ttt gtt gat acg cac cgc 192Asp Ala Ser Asn Met Leu Ile Met Pro Gly Phe Val Asp Thr His Arg 50 55 60cac gta tgg gaa tct gtg atc cgc gga atc gga gcc gac tgg tcc ctg 240His Val Trp Glu Ser Val Ile Arg Gly Ile Gly Ala Asp Trp Ser Leu65 70 75 80caa acc tat tta aac cgg att tat tat ggg aat tat ggg tcg atg aga 288Gln Thr Tyr Leu Asn Arg Ile Tyr Tyr Gly Asn Tyr Gly Ser Met Arg 85 90 95aga cct gag gat gat tgg gcc gcg aac tat tta gga gcg ctt gag gcg 336Arg Pro Glu Asp Asp Trp Ala Ala Asn Tyr Leu Gly Ala Leu Glu Ala 100 105 110ctc gat gca ggc gtc acg act ttg ctg gac tgg acg atg att gag tcg 384Leu Asp Ala Gly Val Thr Thr Leu Leu Asp Trp Thr Met Ile Glu Ser 115 120 125cct gat cat acg gac cag ctg att gcc gga ctg aaa gac gcc ggc att 432Pro Asp His Thr Asp Gln Leu Ile Ala Gly Leu Lys Asp Ala Gly Ile 130 135 140cgc gca gta ttc gcc ttt gga aca tct ggt gac gcc gaa tat tgg gac 480Arg Ala Val Phe Ala Phe Gly Thr Ser Gly Asp Ala Glu Tyr Trp Asp145 150 155 160aga gac agc gat ctt tcg aat atg gac gaa gcg aaa cgg gta aaa aaa 528Arg Asp Ser Asp Leu Ser Asn Met Asp Glu Ala Lys Arg Val Lys Lys 165 170 175gca tat ttt caa tcg aag gat cag ctt ctg acg atg ggg ctt gca att 576Ala Tyr Phe Gln Ser Lys Asp Gln Leu Leu Thr Met Gly Leu Ala Ile 180 185 190cga ggt ccg gag ttt tct tca tgg aag aca tct gtc ttt gaa atc gaa 624Arg Gly Pro Glu Phe Ser Ser Trp Lys Thr Ser Val Phe Glu Ile Glu 195 200 205acg gcc cgg gaa ctg gat gcc tta tgc agc atg cat att gct ttt ggc 672Thr Ala Arg Glu Leu Asp Ala Leu Cys Ser Met His Ile Ala Phe Gly 210 215 220aac tgg gga gcg aaa gat cgt tcg att gaa aag ctg aat caa gcc ggg 720Asn Trp Gly Ala Lys Asp Arg Ser Ile Glu Lys Leu Asn Gln Ala Gly225 230 235 240ctg ctt ggt ccc gat tta aac atg gtg cat gcc aat gcg atg agt gaa 768Leu Leu Gly Pro Asp Leu Asn Met Val His Ala Asn Ala Met Ser Glu 245 250 255gaa gaa atg aaa atg ctt gcg gct cac gga agc tcg ata tct gtg aca 816Glu Glu Met Lys Met Leu Ala Ala His Gly Ser Ser Ile Ser Val Thr 260 265 270cca gaa att gag atg atg atg ggg cac ggc tat cct gtt aca ggg ctc 864Pro Glu Ile Glu Met Met Met Gly His Gly Tyr Pro Val Thr Gly Leu 275 280 285tct cta aaa aac ggg gtt ctt act gcg ctt ggg gtt gat gtt gtc aca 912Ser Leu Lys Asn Gly Val Leu Thr Ala Leu Gly Val Asp Val Val Thr 290 295 300tcg acc ggg gga gac ctt ttc gca cag atg aaa ttt gct ctt caa gca 960Ser Thr Gly Gly Asp Leu Phe Ala Gln Met Lys Phe Ala Leu Gln Ala305 310 315 320gaa cgc gca aag cac aat gat gaa ctg ctg aga gaa gga att atg ccg 1008Glu Arg Ala Lys His Asn Asp Glu Leu Leu Arg Glu Gly Ile Met Pro 325 330 335ggg ccc gaa ttg agc ata tcg gcc gaa caa atc ctg aaa gcg gct acc 1056Gly Pro Glu Leu Ser Ile Ser Ala Glu Gln Ile Leu Lys Ala Ala Thr 340 345 350att gac ggg gcg cgc gcc cta aag ctc gat cat aaa atc ggc tcg ctt 1104Ile Asp Gly Ala Arg Ala Leu Lys Leu Asp His Lys Ile Gly Ser Leu 355 360 365tcc cca ggc aaa gaa gct gac ttt atc atg gtg aaa acg gac agc gtc 1152Ser Pro Gly Lys Glu Ala Asp Phe Ile Met Val Lys Thr Asp Ser Val 370 375 380aat ttg ctc cct ttc act gat gcg gcg ggg gcc atc gtc caa tgc acg 1200Asn Leu Leu Pro Phe Thr Asp Ala Ala Gly Ala Ile Val Gln Cys Thr385 390 395 400cac acc gga aat gta gat tcc gta tat gtc gca gga aaa gcg gtg aaa 1248His Thr Gly Asn Val Asp Ser Val Tyr Val Ala Gly Lys Ala Val Lys 405 410 415cga aac ggc cgg ctg gtt gat gca gat ctt gaa aag gtg aga caa aaa 1296Arg Asn Gly Arg Leu Val Asp Ala Asp Leu Glu Lys Val Arg Gln Lys 420 425 430gcg ctg gaa gca aga gat cat att tta ttc cga tcc cct gag ggc gcc 1344Ala Leu Glu Ala Arg Asp His Ile Leu Phe Arg Ser Pro Glu Gly Ala 435 440 445ttt taa 1350Phe 48449PRTBacillus licheniformis DSM 13 48Met Gln Lys Thr Leu Phe Gln Asn Ala Thr Ile Val Thr Leu Asp Pro1 5 10 15Gln Leu His Thr Leu Glu Lys Gly Asp Met Leu Ile Glu Gly Ser Val 20 25 30Ile Leu Asp Val Ala Glu Ser Ile Glu Ala Gly Asp Ala Glu Ile Val 35 40 45Asp Ala Ser Asn Met Leu Ile Met Pro Gly Phe Val Asp Thr His Arg 50 55 60His Val Trp Glu Ser Val Ile Arg Gly Ile Gly Ala Asp Trp Ser Leu65 70 75 80Gln Thr Tyr Leu Asn Arg Ile Tyr Tyr Gly Asn Tyr Gly Ser Met Arg 85 90 95Arg Pro Glu Asp Asp Trp Ala Ala Asn Tyr Leu Gly Ala Leu Glu Ala 100 105 110Leu Asp Ala Gly Val Thr Thr Leu Leu Asp Trp Thr Met Ile Glu Ser 115 120 125Pro Asp His Thr Asp Gln Leu Ile Ala Gly Leu Lys Asp Ala Gly Ile 130 135 140Arg Ala Val Phe Ala Phe Gly Thr Ser Gly Asp Ala Glu Tyr Trp Asp145 150 155 160Arg Asp Ser Asp Leu Ser Asn Met Asp Glu Ala Lys Arg Val Lys Lys 165 170 175Ala Tyr Phe Gln Ser Lys Asp Gln Leu Leu Thr Met Gly Leu Ala Ile 180 185 190Arg Gly Pro Glu Phe Ser Ser Trp Lys Thr Ser Val Phe Glu Ile Glu 195 200 205Thr Ala Arg Glu Leu Asp Ala Leu Cys Ser Met His Ile Ala Phe Gly 210 215 220Asn Trp Gly Ala Lys Asp Arg Ser Ile Glu Lys Leu Asn Gln Ala Gly225 230 235 240Leu Leu Gly Pro Asp Leu Asn Met Val His Ala Asn Ala Met Ser Glu 245

250 255Glu Glu Met Lys Met Leu Ala Ala His Gly Ser Ser Ile Ser Val Thr 260 265 270Pro Glu Ile Glu Met Met Met Gly His Gly Tyr Pro Val Thr Gly Leu 275 280 285Ser Leu Lys Asn Gly Val Leu Thr Ala Leu Gly Val Asp Val Val Thr 290 295 300Ser Thr Gly Gly Asp Leu Phe Ala Gln Met Lys Phe Ala Leu Gln Ala305 310 315 320Glu Arg Ala Lys His Asn Asp Glu Leu Leu Arg Glu Gly Ile Met Pro 325 330 335Gly Pro Glu Leu Ser Ile Ser Ala Glu Gln Ile Leu Lys Ala Ala Thr 340 345 350Ile Asp Gly Ala Arg Ala Leu Lys Leu Asp His Lys Ile Gly Ser Leu 355 360 365Ser Pro Gly Lys Glu Ala Asp Phe Ile Met Val Lys Thr Asp Ser Val 370 375 380Asn Leu Leu Pro Phe Thr Asp Ala Ala Gly Ala Ile Val Gln Cys Thr385 390 395 400His Thr Gly Asn Val Asp Ser Val Tyr Val Ala Gly Lys Ala Val Lys 405 410 415Arg Asn Gly Arg Leu Val Asp Ala Asp Leu Glu Lys Val Arg Gln Lys 420 425 430Ala Leu Glu Ala Arg Asp His Ile Leu Phe Arg Ser Pro Glu Gly Ala 435 440 445Phe 491359DNABacillus licheniformis DSM 13gene(1)..(1359)putative protein - putative Na(+)-linked D-alanine glycine permease - BLi00817 49atg gaa ttt att tca cat att gta gat aca atg aac ggt ttt tta tgg 48Met Glu Phe Ile Ser His Ile Val Asp Thr Met Asn Gly Phe Leu Trp1 5 10 15ggg cct ccg ctt ttg att ctc att gtt gga aca ggg att tat ttg act 96Gly Pro Pro Leu Leu Ile Leu Ile Val Gly Thr Gly Ile Tyr Leu Thr 20 25 30ttc agg ctg gct ttt ttg cag gtc aga ctt ttg cct tac tct tta aag 144Phe Arg Leu Ala Phe Leu Gln Val Arg Leu Leu Pro Tyr Ser Leu Lys 35 40 45ctc gcc ttt tca agg cat cag gat aaa tcc tcc gaa ggc gat att tcc 192Leu Ala Phe Ser Arg His Gln Asp Lys Ser Ser Glu Gly Asp Ile Ser 50 55 60cac ttt cag gcg ctg atg acc gcc ctt gcc gct acg gtc ggc aca ggg 240His Phe Gln Ala Leu Met Thr Ala Leu Ala Ala Thr Val Gly Thr Gly65 70 75 80aat atc gtc ggc gtc gcc tca gct gtt att gcc ggc ggg ccg ggc gcc 288Asn Ile Val Gly Val Ala Ser Ala Val Ile Ala Gly Gly Pro Gly Ala 85 90 95gta ttt tgg atg tgg ctc gcc gcc ttc ttc gga atg gcg acg aaa tat 336Val Phe Trp Met Trp Leu Ala Ala Phe Phe Gly Met Ala Thr Lys Tyr 100 105 110gct gaa gcg gtg ctt gct gtc aga ttc cgc gtg aaa gat aaa aac ggc 384Ala Glu Ala Val Leu Ala Val Arg Phe Arg Val Lys Asp Lys Asn Gly 115 120 125gaa atg tca ggc ggt ccg atg tat tat ttg gaa cac ggc ctc aag caa 432Glu Met Ser Gly Gly Pro Met Tyr Tyr Leu Glu His Gly Leu Lys Gln 130 135 140aag tgg ctt ggc gtg tta ttt gcc ata ttc ggg gcg ctt gcc gct ttt 480Lys Trp Leu Gly Val Leu Phe Ala Ile Phe Gly Ala Leu Ala Ala Phe145 150 155 160gga ata ggc aat atg gtt cag tca aat tct gta tcc gga gtc atg agc 528Gly Ile Gly Asn Met Val Gln Ser Asn Ser Val Ser Gly Val Met Ser 165 170 175tcc act ttt tcc gtt cct gcc tgg gta acc ggc att att gtc acg act 576Ser Thr Phe Ser Val Pro Ala Trp Val Thr Gly Ile Ile Val Thr Thr 180 185 190ttt acc gcc ctt gtc att ctc ggg gga att aaa agc atc ggc aga gtc 624Phe Thr Ala Leu Val Ile Leu Gly Gly Ile Lys Ser Ile Gly Arg Val 195 200 205acc tcg ctg ttc gtg ccg gtc atg gct tta ttt tat gtc atc gcc gga 672Thr Ser Leu Phe Val Pro Val Met Ala Leu Phe Tyr Val Ile Ala Gly 210 215 220ttg atc att ctt gtc aca aac ttt gaa ctt gtc ccc ggc gct gtc ggt 720Leu Ile Ile Leu Val Thr Asn Phe Glu Leu Val Pro Gly Ala Val Gly225 230 235 240tta att ttc tct gac gcg ttt acc ggg caa gcc gtg gca gga ggc gcg 768Leu Ile Phe Ser Asp Ala Phe Thr Gly Gln Ala Val Ala Gly Gly Ala 245 250 255atc gga acc gtg atc cgc tgg ggc gta gcg cgg ggc gta ttt tca aat 816Ile Gly Thr Val Ile Arg Trp Gly Val Ala Arg Gly Val Phe Ser Asn 260 265 270gaa gca ggt ctc ggc tcg gcg ccg atc gcc gca gcc gcg gca aaa acc 864Glu Ala Gly Leu Gly Ser Ala Pro Ile Ala Ala Ala Ala Ala Lys Thr 275 280 285gat atg ccg gcc cgc cag gcg ctt gtc tct atg acc cag gtt ttc atc 912Asp Met Pro Ala Arg Gln Ala Leu Val Ser Met Thr Gln Val Phe Ile 290 295 300gac aca ttg att gtc tgt tcg att acg ggg ata tca att gtc atg gga 960Asp Thr Leu Ile Val Cys Ser Ile Thr Gly Ile Ser Ile Val Met Gly305 310 315 320aat atg tat ctg aca gaa aaa acc gat gcc ctg aca tcg atg aca ttc 1008Asn Met Tyr Leu Thr Glu Lys Thr Asp Ala Leu Thr Ser Met Thr Phe 325 330 335gca cac ttt ctt ggt cct gcg gga tcg aca atc gtc gcg atc gga ctg 1056Ala His Phe Leu Gly Pro Ala Gly Ser Thr Ile Val Ala Ile Gly Leu 340 345 350ctg ctg ttc gcc tac tcg acg att ttg ggc tgg tcg tac tac gga gaa 1104Leu Leu Phe Ala Tyr Ser Thr Ile Leu Gly Trp Ser Tyr Tyr Gly Glu 355 360 365aaa tgc ttc acc tac ctg ctg aat gaa aaa tat gcg atc gtc tat cgt 1152Lys Cys Phe Thr Tyr Leu Leu Asn Glu Lys Tyr Ala Ile Val Tyr Arg 370 375 380atc atc ttt gtg atc gcc gtc ttt ttc gga gcg gtc ttt aaa aat gac 1200Ile Ile Phe Val Ile Ala Val Phe Phe Gly Ala Val Phe Lys Asn Asp385 390 395 400ttc gtc tgg gcg ctt gcc gat atg ctg aac ggc ctg atg gca att ccg 1248Phe Val Trp Ala Leu Ala Asp Met Leu Asn Gly Leu Met Ala Ile Pro 405 410 415aac ctg atc ggc ctg ctc ggt ttg aca ggc att gtc gtc tat gaa tcc 1296Asn Leu Ile Gly Leu Leu Gly Leu Thr Gly Ile Val Val Tyr Glu Ser 420 425 430aaa cag atc ctc gcc aaa atc aaa gcc gaa aaa gaa gaa aag caa aac 1344Lys Gln Ile Leu Ala Lys Ile Lys Ala Glu Lys Glu Glu Lys Gln Asn 435 440 445agc gta tct gtt taa 1359Ser Val Ser Val 45050452PRTBacillus licheniformis DSM 13 50Met Glu Phe Ile Ser His Ile Val Asp Thr Met Asn Gly Phe Leu Trp1 5 10 15Gly Pro Pro Leu Leu Ile Leu Ile Val Gly Thr Gly Ile Tyr Leu Thr 20 25 30Phe Arg Leu Ala Phe Leu Gln Val Arg Leu Leu Pro Tyr Ser Leu Lys 35 40 45Leu Ala Phe Ser Arg His Gln Asp Lys Ser Ser Glu Gly Asp Ile Ser 50 55 60His Phe Gln Ala Leu Met Thr Ala Leu Ala Ala Thr Val Gly Thr Gly65 70 75 80Asn Ile Val Gly Val Ala Ser Ala Val Ile Ala Gly Gly Pro Gly Ala 85 90 95Val Phe Trp Met Trp Leu Ala Ala Phe Phe Gly Met Ala Thr Lys Tyr 100 105 110Ala Glu Ala Val Leu Ala Val Arg Phe Arg Val Lys Asp Lys Asn Gly 115 120 125Glu Met Ser Gly Gly Pro Met Tyr Tyr Leu Glu His Gly Leu Lys Gln 130 135 140Lys Trp Leu Gly Val Leu Phe Ala Ile Phe Gly Ala Leu Ala Ala Phe145 150 155 160Gly Ile Gly Asn Met Val Gln Ser Asn Ser Val Ser Gly Val Met Ser 165 170 175Ser Thr Phe Ser Val Pro Ala Trp Val Thr Gly Ile Ile Val Thr Thr 180 185 190Phe Thr Ala Leu Val Ile Leu Gly Gly Ile Lys Ser Ile Gly Arg Val 195 200 205Thr Ser Leu Phe Val Pro Val Met Ala Leu Phe Tyr Val Ile Ala Gly 210 215 220Leu Ile Ile Leu Val Thr Asn Phe Glu Leu Val Pro Gly Ala Val Gly225 230 235 240Leu Ile Phe Ser Asp Ala Phe Thr Gly Gln Ala Val Ala Gly Gly Ala 245 250 255Ile Gly Thr Val Ile Arg Trp Gly Val Ala Arg Gly Val Phe Ser Asn 260 265 270Glu Ala Gly Leu Gly Ser Ala Pro Ile Ala Ala Ala Ala Ala Lys Thr 275 280 285Asp Met Pro Ala Arg Gln Ala Leu Val Ser Met Thr Gln Val Phe Ile 290 295 300Asp Thr Leu Ile Val Cys Ser Ile Thr Gly Ile Ser Ile Val Met Gly305 310 315 320Asn Met Tyr Leu Thr Glu Lys Thr Asp Ala Leu Thr Ser Met Thr Phe 325 330 335Ala His Phe Leu Gly Pro Ala Gly Ser Thr Ile Val Ala Ile Gly Leu 340 345 350Leu Leu Phe Ala Tyr Ser Thr Ile Leu Gly Trp Ser Tyr Tyr Gly Glu 355 360 365Lys Cys Phe Thr Tyr Leu Leu Asn Glu Lys Tyr Ala Ile Val Tyr Arg 370 375 380Ile Ile Phe Val Ile Ala Val Phe Phe Gly Ala Val Phe Lys Asn Asp385 390 395 400Phe Val Trp Ala Leu Ala Asp Met Leu Asn Gly Leu Met Ala Ile Pro 405 410 415Asn Leu Ile Gly Leu Leu Gly Leu Thr Gly Ile Val Val Tyr Glu Ser 420 425 430Lys Gln Ile Leu Ala Lys Ile Lys Ala Glu Lys Glu Glu Lys Gln Asn 435 440 445Ser Val Ser Val 450511758DNABacillus licheniformis DSM 13gene(1)..(1758)ggt - gamma-glutamyltranspeptidase - BLi01364 51atg aga cgg tta gct ttc tta gtt gtt gcg ttt tgc ttg gcg gtt ggt 48Met Arg Arg Leu Ala Phe Leu Val Val Ala Phe Cys Leu Ala Val Gly1 5 10 15tgc ttt ttc agt ccg gtg tcg aaa gca gaa gga gtc atg agc ggg gga 96Cys Phe Phe Ser Pro Val Ser Lys Ala Glu Gly Val Met Ser Gly Gly 20 25 30gat ggt gat aaa gtc gcc gtc ggg aaa gac ggt atg gtg gct acg gcc 144Asp Gly Asp Lys Val Ala Val Gly Lys Asp Gly Met Val Ala Thr Ala 35 40 45cat ccg ctt gca tca aaa atc gga gcc gaa gtt ttg aaa aag ggc ggg 192His Pro Leu Ala Ser Lys Ile Gly Ala Glu Val Leu Lys Lys Gly Gly 50 55 60aat gcc att gat gca gct att gcg atc cag tac gca ctg aat gta aca 240Asn Ala Ile Asp Ala Ala Ile Ala Ile Gln Tyr Ala Leu Asn Val Thr65 70 75 80gag ccg atg atg tct gga atc ggc ggc ggc ggt ttc atg atg gtc tac 288Glu Pro Met Met Ser Gly Ile Gly Gly Gly Gly Phe Met Met Val Tyr 85 90 95gac ggc gag acg aag gaa acg tcg atc atc aac agc agg gag cgt gct 336Asp Gly Glu Thr Lys Glu Thr Ser Ile Ile Asn Ser Arg Glu Arg Ala 100 105 110ccg gag gga gcc aag cct gac atg ttt ttg gat gaa gac gga aag gtg 384Pro Glu Gly Ala Lys Pro Asp Met Phe Leu Asp Glu Asp Gly Lys Val 115 120 125att ccg ttt tca gaa agg tcg aga cac gga aat gcg gtc ggt gtg ccg 432Ile Pro Phe Ser Glu Arg Ser Arg His Gly Asn Ala Val Gly Val Pro 130 135 140gga acg ctg aaa ggg ctt gaa gca gcg cat aag aag tgg gga aca aag 480Gly Thr Leu Lys Gly Leu Glu Ala Ala His Lys Lys Trp Gly Thr Lys145 150 155 160aaa atg gaa gac ttg att tcg cct tct ata aag ctt gca gag gaa ggg 528Lys Met Glu Asp Leu Ile Ser Pro Ser Ile Lys Leu Ala Glu Glu Gly 165 170 175ttc cca atc gac tcg gtt ttg gct gac gcg ata aag gat cat cag gac 576Phe Pro Ile Asp Ser Val Leu Ala Asp Ala Ile Lys Asp His Gln Asp 180 185 190aag ctg agc aaa acg gcc gca aag gat att ttc ctt ccg gac ggt gaa 624Lys Leu Ser Lys Thr Ala Ala Lys Asp Ile Phe Leu Pro Asp Gly Glu 195 200 205cct ttg aaa gaa ggc gat atc ctt gta caa aag gat ttg gcc aaa acg 672Pro Leu Lys Glu Gly Asp Ile Leu Val Gln Lys Asp Leu Ala Lys Thr 210 215 220ttt aaa ttg ata cgg aaa gaa ggg tca aaa gcc ttt tat gat ggg gaa 720Phe Lys Leu Ile Arg Lys Glu Gly Ser Lys Ala Phe Tyr Asp Gly Glu225 230 235 240atc ggc cgg gcg att gca gat gtc gtg cag gat ttc ggg ggt tca atg 768Ile Gly Arg Ala Ile Ala Asp Val Val Gln Asp Phe Gly Gly Ser Met 245 250 255aca cct gat gac ctg tcc cgc tac gag gtc aca aca gat aag ccc atc 816Thr Pro Asp Asp Leu Ser Arg Tyr Glu Val Thr Thr Asp Lys Pro Ile 260 265 270tgg ggc gag tac cac ggc tat gat att gca agc atg cct cct cca agc 864Trp Gly Glu Tyr His Gly Tyr Asp Ile Ala Ser Met Pro Pro Pro Ser 275 280 285tca ggc gga gtg ttc atg ctg cag atg ctg aaa ttg att gat gat ttt 912Ser Gly Gly Val Phe Met Leu Gln Met Leu Lys Leu Ile Asp Asp Phe 290 295 300cat tta tcg caa tac gat cca aag tcg ttt gaa aaa tat cat ctg ctc 960His Leu Ser Gln Tyr Asp Pro Lys Ser Phe Glu Lys Tyr His Leu Leu305 310 315 320gcg gaa acg atg cat ctc tcc tat gcg gac agg gct gca tat gcg ggc 1008Ala Glu Thr Met His Leu Ser Tyr Ala Asp Arg Ala Ala Tyr Ala Gly 325 330 335gat ccg gaa ttt gtc gat gtt ccg ctc aga ggg ctg ctc gat ccc gat 1056Asp Pro Glu Phe Val Asp Val Pro Leu Arg Gly Leu Leu Asp Pro Asp 340 345 350tat att aag gag cgg caa aag ctg atc agc ctt gat tcg atg aac cgc 1104Tyr Ile Lys Glu Arg Gln Lys Leu Ile Ser Leu Asp Ser Met Asn Arg 355 360 365gat gtg aaa gag ggc gat ccg tgg aaa tat gag gaa ggc gag ccg aat 1152Asp Val Lys Glu Gly Asp Pro Trp Lys Tyr Glu Glu Gly Glu Pro Asn 370 375 380tat gag atc gtg cct cag cct gaa gat aaa acg atc ggg gag acg acg 1200Tyr Glu Ile Val Pro Gln Pro Glu Asp Lys Thr Ile Gly Glu Thr Thr385 390 395 400cat ttt acc gtc acg gat cag tgg ggc aat gtt gtc tct tat acg aca 1248His Phe Thr Val Thr Asp Gln Trp Gly Asn Val Val Ser Tyr Thr Thr 405 410 415acg att gaa cag ctt ttt gga acg ggg att tta gtg ccc ggg tac gga 1296Thr Ile Glu Gln Leu Phe Gly Thr Gly Ile Leu Val Pro Gly Tyr Gly 420 425 430ctg ttt ttg aac aat gaa ctg acc gat ttt gat gcg att cca ggc ggc 1344Leu Phe Leu Asn Asn Glu Leu Thr Asp Phe Asp Ala Ile Pro Gly Gly 435 440 445gcc aac gaa gtg cag ccg aat aaa agg ccg ctt tca agc atg acg ccg 1392Ala Asn Glu Val Gln Pro Asn Lys Arg Pro Leu Ser Ser Met Thr Pro 450 455 460acg atc gta ttc aaa gat gaa aaa ccg gtg ctg act gtc ggt tcc cct 1440Thr Ile Val Phe Lys Asp Glu Lys Pro Val Leu Thr Val Gly Ser Pro465 470 475 480ggc gga acg acg atc att gct tcc gtt ttc cag acg atc ttg aat tat 1488Gly Gly Thr Thr Ile Ile Ala Ser Val Phe Gln Thr Ile Leu Asn Tyr 485 490 495ttt gag tac ggc atg agc ctt cag gat gcg att gaa gag ccg agg atc 1536Phe Glu Tyr Gly Met Ser Leu Gln Asp Ala Ile Glu Glu Pro Arg Ile 500 505 510tat acg aac agc ctg act tcg tac cgc tat gaa tcc gga atg ccg gag 1584Tyr Thr Asn Ser Leu Thr Ser Tyr Arg Tyr Glu Ser Gly Met Pro Glu 515 520 525gat gtc aga cgg aag ctc aat gac ttc ggc cac aaa ttc ggc tcg aat 1632Asp Val Arg Arg Lys Leu Asn Asp Phe Gly His Lys Phe Gly Ser Asn 530 535 540ccg gtt gat atc ggc aat gtc caa agc att ttc att gac cgg gag aac 1680Pro Val Asp Ile Gly Asn Val Gln Ser Ile Phe Ile Asp Arg Glu Asn545 550 555 560aaa acg ttt atg ggg gtt gcc gat tca agc aga aac gga acg gcg gtt 1728Lys Thr Phe Met Gly Val Ala Asp Ser Ser Arg Asn Gly Thr Ala Val 565 570 575ggg gtc aac att aag aca tcg gct aaa tag 1758Gly Val Asn Ile Lys Thr Ser Ala Lys 580 58552585PRTBacillus licheniformis DSM 13 52Met Arg Arg Leu Ala Phe Leu Val Val Ala Phe Cys Leu Ala Val Gly1 5 10 15Cys Phe Phe Ser Pro Val Ser Lys Ala Glu Gly Val Met Ser Gly Gly 20 25 30Asp Gly Asp Lys Val Ala Val Gly Lys Asp Gly Met Val Ala Thr Ala 35 40 45His Pro Leu Ala Ser Lys Ile Gly Ala Glu Val Leu Lys Lys Gly Gly 50 55 60Asn Ala Ile Asp Ala Ala Ile Ala Ile Gln Tyr Ala Leu Asn Val Thr65 70 75 80Glu Pro Met Met Ser Gly Ile Gly Gly Gly Gly Phe Met Met Val Tyr 85 90 95Asp Gly Glu Thr Lys Glu Thr Ser Ile Ile Asn Ser Arg Glu Arg Ala

100 105 110Pro Glu Gly Ala Lys Pro Asp Met Phe Leu Asp Glu Asp Gly Lys Val 115 120 125Ile Pro Phe Ser Glu Arg Ser Arg His Gly Asn Ala Val Gly Val Pro 130 135 140Gly Thr Leu Lys Gly Leu Glu Ala Ala His Lys Lys Trp Gly Thr Lys145 150 155 160Lys Met Glu Asp Leu Ile Ser Pro Ser Ile Lys Leu Ala Glu Glu Gly 165 170 175Phe Pro Ile Asp Ser Val Leu Ala Asp Ala Ile Lys Asp His Gln Asp 180 185 190Lys Leu Ser Lys Thr Ala Ala Lys Asp Ile Phe Leu Pro Asp Gly Glu 195 200 205Pro Leu Lys Glu Gly Asp Ile Leu Val Gln Lys Asp Leu Ala Lys Thr 210 215 220Phe Lys Leu Ile Arg Lys Glu Gly Ser Lys Ala Phe Tyr Asp Gly Glu225 230 235 240Ile Gly Arg Ala Ile Ala Asp Val Val Gln Asp Phe Gly Gly Ser Met 245 250 255Thr Pro Asp Asp Leu Ser Arg Tyr Glu Val Thr Thr Asp Lys Pro Ile 260 265 270Trp Gly Glu Tyr His Gly Tyr Asp Ile Ala Ser Met Pro Pro Pro Ser 275 280 285Ser Gly Gly Val Phe Met Leu Gln Met Leu Lys Leu Ile Asp Asp Phe 290 295 300His Leu Ser Gln Tyr Asp Pro Lys Ser Phe Glu Lys Tyr His Leu Leu305 310 315 320Ala Glu Thr Met His Leu Ser Tyr Ala Asp Arg Ala Ala Tyr Ala Gly 325 330 335Asp Pro Glu Phe Val Asp Val Pro Leu Arg Gly Leu Leu Asp Pro Asp 340 345 350Tyr Ile Lys Glu Arg Gln Lys Leu Ile Ser Leu Asp Ser Met Asn Arg 355 360 365Asp Val Lys Glu Gly Asp Pro Trp Lys Tyr Glu Glu Gly Glu Pro Asn 370 375 380Tyr Glu Ile Val Pro Gln Pro Glu Asp Lys Thr Ile Gly Glu Thr Thr385 390 395 400His Phe Thr Val Thr Asp Gln Trp Gly Asn Val Val Ser Tyr Thr Thr 405 410 415Thr Ile Glu Gln Leu Phe Gly Thr Gly Ile Leu Val Pro Gly Tyr Gly 420 425 430Leu Phe Leu Asn Asn Glu Leu Thr Asp Phe Asp Ala Ile Pro Gly Gly 435 440 445Ala Asn Glu Val Gln Pro Asn Lys Arg Pro Leu Ser Ser Met Thr Pro 450 455 460Thr Ile Val Phe Lys Asp Glu Lys Pro Val Leu Thr Val Gly Ser Pro465 470 475 480Gly Gly Thr Thr Ile Ile Ala Ser Val Phe Gln Thr Ile Leu Asn Tyr 485 490 495Phe Glu Tyr Gly Met Ser Leu Gln Asp Ala Ile Glu Glu Pro Arg Ile 500 505 510Tyr Thr Asn Ser Leu Thr Ser Tyr Arg Tyr Glu Ser Gly Met Pro Glu 515 520 525Asp Val Arg Arg Lys Leu Asn Asp Phe Gly His Lys Phe Gly Ser Asn 530 535 540Pro Val Asp Ile Gly Asn Val Gln Ser Ile Phe Ile Asp Arg Glu Asn545 550 555 560Lys Thr Phe Met Gly Val Ala Asp Ser Ser Arg Asn Gly Thr Ala Val 565 570 575Gly Val Asn Ile Lys Thr Ser Ala Lys 580 58553942DNABacillus licheniformis DSM 13gene(1)..(942)hypothetical protein - unknown function - BLi04116 53atg agg gtc ttg aaa aat gaa ctt tac agg ctg atg gtg acg aaa agt 48Met Arg Val Leu Lys Asn Glu Leu Tyr Arg Leu Met Val Thr Lys Ser1 5 10 15acc tgg att gtg tta agc ttg ctg ctt gtc atg aca atc gct gtt gca 96Thr Trp Ile Val Leu Ser Leu Leu Leu Val Met Thr Ile Ala Val Ala 20 25 30tgg atg gtc agc aat ggc gaa aag gag aag gag aca ggt aac tgg aaa 144Trp Met Val Ser Asn Gly Glu Lys Glu Lys Glu Thr Gly Asn Trp Lys 35 40 45gag caa tta acc gtt caa aac gct cag tat gaa aga gaa atg aga gag 192Glu Gln Leu Thr Val Gln Asn Ala Gln Tyr Glu Arg Glu Met Arg Glu 50 55 60ctg agc cca gcg gtt ccc aaa tac caa ttt tta aaa gaa gag atc gcg 240Leu Ser Pro Ala Val Pro Lys Tyr Gln Phe Leu Lys Glu Glu Ile Ala65 70 75 80gtc aat caa tac cgg ctt gag cat aat ttg ccg cct tct gcg aaa tac 288Val Asn Gln Tyr Arg Leu Glu His Asn Leu Pro Pro Ser Ala Lys Tyr 85 90 95aat gtt tgg acg atg ctg aag gag tta aaa ccg atc aca aca tta atc 336Asn Val Trp Thr Met Leu Lys Glu Leu Lys Pro Ile Thr Thr Leu Ile 100 105 110gcg ttg att gca atc gtg ctg gca gcg aac tcc atc gcg ctt gag cac 384Ala Leu Ile Ala Ile Val Leu Ala Ala Asn Ser Ile Ala Leu Glu His 115 120 125agc aaa gga act att aaa ttt gcg att gca aca ccg gtc aag cgt tgg 432Ser Lys Gly Thr Ile Lys Phe Ala Ile Ala Thr Pro Val Lys Arg Trp 130 135 140cat tat cta ttg ggg aaa tac ttg tcg atc ttg ctc aac act gta ttt 480His Tyr Leu Leu Gly Lys Tyr Leu Ser Ile Leu Leu Asn Thr Val Phe145 150 155 160atg ttt gct gcg aca ctt tta ttt gcg ttt gtt tta ggg tat gcc ctg 528Met Phe Ala Ala Thr Leu Leu Phe Ala Phe Val Leu Gly Tyr Ala Leu 165 170 175cta ggt ttg gag gga agc caa tac tat ttg tcc tat cga agc ggc gaa 576Leu Gly Leu Glu Gly Ser Gln Tyr Tyr Leu Ser Tyr Arg Ser Gly Glu 180 185 190gtg ata aag atg tca atg ctg aag ttt tta gcc tta gac tat gga gcc 624Val Ile Lys Met Ser Met Leu Lys Phe Leu Ala Leu Asp Tyr Gly Ala 195 200 205gct tta ttg aat att att gtg ctg gcc acg ctg gcc ttt atg att tcg 672Ala Leu Leu Asn Ile Ile Val Leu Ala Thr Leu Ala Phe Met Ile Ser 210 215 220gtc atc cta aga agc gcg gtc gtc tct gtc ggt ctt tcg ctg ttc gtc 720Val Ile Leu Arg Ser Ala Val Val Ser Val Gly Leu Ser Leu Phe Val225 230 235 240ttt ttt aca ggg tct gcg att act caa ttt tta gcg gca aaa ttt gat 768Phe Phe Thr Gly Ser Ala Ile Thr Gln Phe Leu Ala Ala Lys Phe Asp 245 250 255tgg acc aag tat acg atc ttt gca aat tca gat ctc agt caa tac att 816Trp Thr Lys Tyr Thr Ile Phe Ala Asn Ser Asp Leu Ser Gln Tyr Ile 260 265 270gat ggg gag ccg ttt att caa gat atg act ctt tca ttt tca gcg gct 864Asp Gly Glu Pro Phe Ile Gln Asp Met Thr Leu Ser Phe Ser Ala Ala 275 280 285gtg att gct gtt tat ttt att ctt ttt ctg gct gtc tcc ttt tgg gtt 912Val Ile Ala Val Tyr Phe Ile Leu Phe Leu Ala Val Ser Phe Trp Val 290 295 300ttt caa aag cgg gat att gtc acg tca taa 942Phe Gln Lys Arg Asp Ile Val Thr Ser305 31054313PRTBacillus licheniformis DSM 13 54Met Arg Val Leu Lys Asn Glu Leu Tyr Arg Leu Met Val Thr Lys Ser1 5 10 15Thr Trp Ile Val Leu Ser Leu Leu Leu Val Met Thr Ile Ala Val Ala 20 25 30Trp Met Val Ser Asn Gly Glu Lys Glu Lys Glu Thr Gly Asn Trp Lys 35 40 45Glu Gln Leu Thr Val Gln Asn Ala Gln Tyr Glu Arg Glu Met Arg Glu 50 55 60Leu Ser Pro Ala Val Pro Lys Tyr Gln Phe Leu Lys Glu Glu Ile Ala65 70 75 80Val Asn Gln Tyr Arg Leu Glu His Asn Leu Pro Pro Ser Ala Lys Tyr 85 90 95Asn Val Trp Thr Met Leu Lys Glu Leu Lys Pro Ile Thr Thr Leu Ile 100 105 110Ala Leu Ile Ala Ile Val Leu Ala Ala Asn Ser Ile Ala Leu Glu His 115 120 125Ser Lys Gly Thr Ile Lys Phe Ala Ile Ala Thr Pro Val Lys Arg Trp 130 135 140His Tyr Leu Leu Gly Lys Tyr Leu Ser Ile Leu Leu Asn Thr Val Phe145 150 155 160Met Phe Ala Ala Thr Leu Leu Phe Ala Phe Val Leu Gly Tyr Ala Leu 165 170 175Leu Gly Leu Glu Gly Ser Gln Tyr Tyr Leu Ser Tyr Arg Ser Gly Glu 180 185 190Val Ile Lys Met Ser Met Leu Lys Phe Leu Ala Leu Asp Tyr Gly Ala 195 200 205Ala Leu Leu Asn Ile Ile Val Leu Ala Thr Leu Ala Phe Met Ile Ser 210 215 220Val Ile Leu Arg Ser Ala Val Val Ser Val Gly Leu Ser Leu Phe Val225 230 235 240Phe Phe Thr Gly Ser Ala Ile Thr Gln Phe Leu Ala Ala Lys Phe Asp 245 250 255Trp Thr Lys Tyr Thr Ile Phe Ala Asn Ser Asp Leu Ser Gln Tyr Ile 260 265 270Asp Gly Glu Pro Phe Ile Gln Asp Met Thr Leu Ser Phe Ser Ala Ala 275 280 285Val Ile Ala Val Tyr Phe Ile Leu Phe Leu Ala Val Ser Phe Trp Val 290 295 300Phe Gln Lys Arg Asp Ile Val Thr Ser305 31055903DNABacillus licheniformis DSM 13gene(1)..(903)putative protein - putative ABC transporter ATP-binding protein - BLi04117 55atg agt caa gtc gaa ttt gaa ggt gta agt aaa cga ata aaa ggc aga 48Met Ser Gln Val Glu Phe Glu Gly Val Ser Lys Arg Ile Lys Gly Arg1 5 10 15cca att gtc caa aat atc aca ttt caa att gcc cca ggt aca att ttt 96Pro Ile Val Gln Asn Ile Thr Phe Gln Ile Ala Pro Gly Thr Ile Phe 20 25 30ggg ctg ctc ggg cca aac ggc gct ggc aag aca aca ctt atc aaa atg 144Gly Leu Leu Gly Pro Asn Gly Ala Gly Lys Thr Thr Leu Ile Lys Met 35 40 45att gtc ggg atg gca aag ccg aca tca gga gat atc cgc atc gac ggc 192Ile Val Gly Met Ala Lys Pro Thr Ser Gly Asp Ile Arg Ile Asp Gly 50 55 60tat tca gtt aaa agc aat tac gag gaa gcg gca gcc cga gtc ggt tct 240Tyr Ser Val Lys Ser Asn Tyr Glu Glu Ala Ala Ala Arg Val Gly Ser65 70 75 80gtt gtt gaa aac cca tcc ttt tat gag cac tta aca gga tac caa aac 288Val Val Glu Asn Pro Ser Phe Tyr Glu His Leu Thr Gly Tyr Gln Asn 85 90 95ctt aaa tat ctc ggc gga ttc cac agc cac gtg tca aag gag cgc ata 336Leu Lys Tyr Leu Gly Gly Phe His Ser His Val Ser Lys Glu Arg Ile 100 105 110gaa gag atc gtt cag ctt gtt gat ttg aca gga agt att cat aaa cca 384Glu Glu Ile Val Gln Leu Val Asp Leu Thr Gly Ser Ile His Lys Pro 115 120 125gtt aaa acg tat tca tta ggc atg aaa cag cgt ttg ggc ctt gcc gtc 432Val Lys Thr Tyr Ser Leu Gly Met Lys Gln Arg Leu Gly Leu Ala Val 130 135 140gcg ctc ttg cat gat ccg gaa ttt ctc att ctc gat gaa ccg aca aac 480Ala Leu Leu His Asp Pro Glu Phe Leu Ile Leu Asp Glu Pro Thr Asn145 150 155 160ggc ctt gat cct cag gga atc att gat ttg cgc gaa cac ctt cag tac 528Gly Leu Asp Pro Gln Gly Ile Ile Asp Leu Arg Glu His Leu Gln Tyr 165 170 175ttg gcg aaa acc ttc aac aaa acg att ttg att tcg agt cat ctt ctg 576Leu Ala Lys Thr Phe Asn Lys Thr Ile Leu Ile Ser Ser His Leu Leu 180 185 190tct gag gtt gag atg att tgt gat gaa tac ggc gtc atg aaa aac gga 624Ser Glu Val Glu Met Ile Cys Asp Glu Tyr Gly Val Met Lys Asn Gly 195 200 205gaa ctc ctg caa att aag agc aat cac cgc gat acc gat acg gtt cgt 672Glu Leu Leu Gln Ile Lys Ser Asn His Arg Asp Thr Asp Thr Val Arg 210 215 220tat cgg ctt aca tta aac ggc cac gcc gat gaa gcg gct gac ctg ttg 720Tyr Arg Leu Thr Leu Asn Gly His Ala Asp Glu Ala Ala Asp Leu Leu225 230 235 240aat gag tac cag tat gca ggc ggt ctc acg gaa gat aaa aat gag att 768Asn Glu Tyr Gln Tyr Ala Gly Gly Leu Thr Glu Asp Lys Asn Glu Ile 245 250 255tat gtc ctt tgc atg gaa gaa gac att atg aaa gtc gtt aat ctg tta 816Tyr Val Leu Cys Met Glu Glu Asp Ile Met Lys Val Val Asn Leu Leu 260 265 270atg gag aac aaa ata aga gtt ctg cat atg aag cag gaa aaa cag tcg 864Met Glu Asn Lys Ile Arg Val Leu His Met Lys Gln Glu Lys Gln Ser 275 280 285ata gaa caa agc ttt ctg gaa ttg atc aat aag ggg tga 903Ile Glu Gln Ser Phe Leu Glu Leu Ile Asn Lys Gly 290 295 30056300PRTBacillus licheniformis DSM 13 56Met Ser Gln Val Glu Phe Glu Gly Val Ser Lys Arg Ile Lys Gly Arg1 5 10 15Pro Ile Val Gln Asn Ile Thr Phe Gln Ile Ala Pro Gly Thr Ile Phe 20 25 30Gly Leu Leu Gly Pro Asn Gly Ala Gly Lys Thr Thr Leu Ile Lys Met 35 40 45Ile Val Gly Met Ala Lys Pro Thr Ser Gly Asp Ile Arg Ile Asp Gly 50 55 60Tyr Ser Val Lys Ser Asn Tyr Glu Glu Ala Ala Ala Arg Val Gly Ser65 70 75 80Val Val Glu Asn Pro Ser Phe Tyr Glu His Leu Thr Gly Tyr Gln Asn 85 90 95Leu Lys Tyr Leu Gly Gly Phe His Ser His Val Ser Lys Glu Arg Ile 100 105 110Glu Glu Ile Val Gln Leu Val Asp Leu Thr Gly Ser Ile His Lys Pro 115 120 125Val Lys Thr Tyr Ser Leu Gly Met Lys Gln Arg Leu Gly Leu Ala Val 130 135 140Ala Leu Leu His Asp Pro Glu Phe Leu Ile Leu Asp Glu Pro Thr Asn145 150 155 160Gly Leu Asp Pro Gln Gly Ile Ile Asp Leu Arg Glu His Leu Gln Tyr 165 170 175Leu Ala Lys Thr Phe Asn Lys Thr Ile Leu Ile Ser Ser His Leu Leu 180 185 190Ser Glu Val Glu Met Ile Cys Asp Glu Tyr Gly Val Met Lys Asn Gly 195 200 205Glu Leu Leu Gln Ile Lys Ser Asn His Arg Asp Thr Asp Thr Val Arg 210 215 220Tyr Arg Leu Thr Leu Asn Gly His Ala Asp Glu Ala Ala Asp Leu Leu225 230 235 240Asn Glu Tyr Gln Tyr Ala Gly Gly Leu Thr Glu Asp Lys Asn Glu Ile 245 250 255Tyr Val Leu Cys Met Glu Glu Asp Ile Met Lys Val Val Asn Leu Leu 260 265 270Met Glu Asn Lys Ile Arg Val Leu His Met Lys Gln Glu Lys Gln Ser 275 280 285Ile Glu Gln Ser Phe Leu Glu Leu Ile Asn Lys Gly 290 295 300571200DNABacillus licheniformis DSM 13gene(1)..(1200)yciC - unknown function - similar to unknown function proteins - BLi00765 57atg agt aaa aag att cct gtg aca gta tta agc gga tac ctg ggg gcg 48Met Ser Lys Lys Ile Pro Val Thr Val Leu Ser Gly Tyr Leu Gly Ala1 5 10 15ggt aaa acg acg ctg ctg aat cac ata ttg caa aac cgc caa ggg ctg 96Gly Lys Thr Thr Leu Leu Asn His Ile Leu Gln Asn Arg Gln Gly Leu 20 25 30aaa gtc gcc gtt att gtt aat gac atg agc gaa gtc aac att gat gca 144Lys Val Ala Val Ile Val Asn Asp Met Ser Glu Val Asn Ile Asp Ala 35 40 45ggg ctt gtc aag caa ggc ggc ctg tcg agg acg gat gaa aag ctg gtc 192Gly Leu Val Lys Gln Gly Gly Leu Ser Arg Thr Asp Glu Lys Leu Val 50 55 60gaa atg tca aac ggc tgc att tgc tgc aca ctc aga gag gat ttg ctg 240Glu Met Ser Asn Gly Cys Ile Cys Cys Thr Leu Arg Glu Asp Leu Leu65 70 75 80gtt gaa gtc gaa aag ctg gcg aag cag gga aat att gat tat atc gtg 288Val Glu Val Glu Lys Leu Ala Lys Gln Gly Asn Ile Asp Tyr Ile Val 85 90 95atc gaa tcg acc ggt atc agc gaa ccg atc ccg gtc gct cag acg ttc 336Ile Glu Ser Thr Gly Ile Ser Glu Pro Ile Pro Val Ala Gln Thr Phe 100 105 110tct tac atc gac gaa gaa ttg ggt att gat ttg acc cgg ttc tgc cgc 384Ser Tyr Ile Asp Glu Glu Leu Gly Ile Asp Leu Thr Arg Phe Cys Arg 115 120 125ttg gac aca atg gtg acg gtt gtc gac gcc aat cgt ttt tgg cac gat 432Leu Asp Thr Met Val Thr Val Val Asp Ala Asn Arg Phe Trp His Asp 130 135 140ttt cag tcg ggg gac agc ctg ctg gac cgg aaa gag gct gtc ggc gaa 480Phe Gln Ser Gly Asp Ser Leu Leu Asp Arg Lys Glu Ala Val Gly Glu145 150 155 160gag gat gaa cgc gac atc gcc gat ctg ctc atc gat caa atc gag ttc 528Glu Asp Glu Arg Asp Ile Ala Asp Leu Leu Ile Asp Gln Ile Glu Phe 165 170 175tgc gat gtg ctg att ttg aat aaa tgc gac ctt gtc tcc gag gaa gat 576Cys Asp Val Leu Ile Leu Asn Lys Cys Asp Leu Val Ser Glu Glu Asp 180 185 190ctg caa aag ctt gag aaa gtg ctg aaa act ttg cag cct aaa gca aac 624Leu Gln Lys Leu Glu Lys Val Leu Lys Thr Leu Gln Pro Lys Ala Asn 195 200

205atc atc aga gcg gtc aaa gga gag gtc gat ccg aag gcc gtc tta aac 672Ile Ile Arg Ala Val Lys Gly Glu Val Asp Pro Lys Ala Val Leu Asn 210 215 220acc ggt ctc ttc aat ttt gac gaa gcg agc ggt tca gca ggc tgg atc 720Thr Gly Leu Phe Asn Phe Asp Glu Ala Ser Gly Ser Ala Gly Trp Ile225 230 235 240agg gaa tta aac caa gga cac cac cag cat acg cct gag act gaa gaa 768Arg Glu Leu Asn Gln Gly His His Gln His Thr Pro Glu Thr Glu Glu 245 250 255tac gga ata tct tcg ttt gta tac gaa gcg cgc cgt cct ttt cat aca 816Tyr Gly Ile Ser Ser Phe Val Tyr Glu Ala Arg Arg Pro Phe His Thr 260 265 270gag cgg ttt tat aac tgg att cat tcg ctg ccg gaa aac gtc gtc agg 864Glu Arg Phe Tyr Asn Trp Ile His Ser Leu Pro Glu Asn Val Val Arg 275 280 285gcg aaa gga ata gcc tgg tgt gcg acg cgc aac acc ctt gcc ctt ttg 912Ala Lys Gly Ile Ala Trp Cys Ala Thr Arg Asn Thr Leu Ala Leu Leu 290 295 300atg tcg cag gcg ggt cct tcc gtg tcg ctg gaa ccg atc tct tat tgg 960Met Ser Gln Ala Gly Pro Ser Val Ser Leu Glu Pro Ile Ser Tyr Trp305 310 315 320gtg gcc gca ctg tcc aaa ccg gaa cag gaa caa att ctc aga cag aac 1008Val Ala Ala Leu Ser Lys Pro Glu Gln Glu Gln Ile Leu Arg Gln Asn 325 330 335cct gag ctt tta aaa gaa tgg aat gaa gaa ttc gga gac cgc cat aca 1056Pro Glu Leu Leu Lys Glu Trp Asn Glu Glu Phe Gly Asp Arg His Thr 340 345 350aag ctc gtc ttt atc gga ttg gat ctg tcg ccg tct gaa att acc gca 1104Lys Leu Val Phe Ile Gly Leu Asp Leu Ser Pro Ser Glu Ile Thr Ala 355 360 365gaa gcg gat cga tgc ctt tta aca gat tcg gag atg gcg gcc gac tgg 1152Glu Ala Asp Arg Cys Leu Leu Thr Asp Ser Glu Met Ala Ala Asp Trp 370 375 380tcg ctt tta tct gac ccg ttc gat tgg cag att gaa aga gct aga taa 1200Ser Leu Leu Ser Asp Pro Phe Asp Trp Gln Ile Glu Arg Ala Arg385 390 39558399PRTBacillus licheniformis DSM 13 58Met Ser Lys Lys Ile Pro Val Thr Val Leu Ser Gly Tyr Leu Gly Ala1 5 10 15Gly Lys Thr Thr Leu Leu Asn His Ile Leu Gln Asn Arg Gln Gly Leu 20 25 30Lys Val Ala Val Ile Val Asn Asp Met Ser Glu Val Asn Ile Asp Ala 35 40 45Gly Leu Val Lys Gln Gly Gly Leu Ser Arg Thr Asp Glu Lys Leu Val 50 55 60Glu Met Ser Asn Gly Cys Ile Cys Cys Thr Leu Arg Glu Asp Leu Leu65 70 75 80Val Glu Val Glu Lys Leu Ala Lys Gln Gly Asn Ile Asp Tyr Ile Val 85 90 95Ile Glu Ser Thr Gly Ile Ser Glu Pro Ile Pro Val Ala Gln Thr Phe 100 105 110Ser Tyr Ile Asp Glu Glu Leu Gly Ile Asp Leu Thr Arg Phe Cys Arg 115 120 125Leu Asp Thr Met Val Thr Val Val Asp Ala Asn Arg Phe Trp His Asp 130 135 140Phe Gln Ser Gly Asp Ser Leu Leu Asp Arg Lys Glu Ala Val Gly Glu145 150 155 160Glu Asp Glu Arg Asp Ile Ala Asp Leu Leu Ile Asp Gln Ile Glu Phe 165 170 175Cys Asp Val Leu Ile Leu Asn Lys Cys Asp Leu Val Ser Glu Glu Asp 180 185 190Leu Gln Lys Leu Glu Lys Val Leu Lys Thr Leu Gln Pro Lys Ala Asn 195 200 205Ile Ile Arg Ala Val Lys Gly Glu Val Asp Pro Lys Ala Val Leu Asn 210 215 220Thr Gly Leu Phe Asn Phe Asp Glu Ala Ser Gly Ser Ala Gly Trp Ile225 230 235 240Arg Glu Leu Asn Gln Gly His His Gln His Thr Pro Glu Thr Glu Glu 245 250 255Tyr Gly Ile Ser Ser Phe Val Tyr Glu Ala Arg Arg Pro Phe His Thr 260 265 270Glu Arg Phe Tyr Asn Trp Ile His Ser Leu Pro Glu Asn Val Val Arg 275 280 285Ala Lys Gly Ile Ala Trp Cys Ala Thr Arg Asn Thr Leu Ala Leu Leu 290 295 300Met Ser Gln Ala Gly Pro Ser Val Ser Leu Glu Pro Ile Ser Tyr Trp305 310 315 320Val Ala Ala Leu Ser Lys Pro Glu Gln Glu Gln Ile Leu Arg Gln Asn 325 330 335Pro Glu Leu Leu Lys Glu Trp Asn Glu Glu Phe Gly Asp Arg His Thr 340 345 350Lys Leu Val Phe Ile Gly Leu Asp Leu Ser Pro Ser Glu Ile Thr Ala 355 360 365Glu Ala Asp Arg Cys Leu Leu Thr Asp Ser Glu Met Ala Ala Asp Trp 370 375 380Ser Leu Leu Ser Asp Pro Phe Asp Trp Gln Ile Glu Arg Ala Arg385 390 395591092DNABacillus licheniformis DSM 13gene(1)..(1092)citA - citrate synthase I (minor) - BLi01010 59atg gtt tat cat ggg tta aaa ggg att act tgt gtt gaa aca gcc atc 48Met Val Tyr His Gly Leu Lys Gly Ile Thr Cys Val Glu Thr Ala Ile1 5 10 15agc cgc att gac ggt gaa aaa gga aag ctg atc tac aga ggg tac aat 96Ser Arg Ile Asp Gly Glu Lys Gly Lys Leu Ile Tyr Arg Gly Tyr Asn 20 25 30gct gat gaa ctg gcg ctt tcg tcc aca ttt gaa gaa gtc gct tac ctg 144Ala Asp Glu Leu Ala Leu Ser Ser Thr Phe Glu Glu Val Ala Tyr Leu 35 40 45att ctc aac ggg gca ttt ccg acg gaa gct gaa ctc agc aga ttc aat 192Ile Leu Asn Gly Ala Phe Pro Thr Glu Ala Glu Leu Ser Arg Phe Asn 50 55 60gag ctg ctc gct tca ttt cgt gtg ctt cca gaa gca tgc gag cga ttg 240Glu Leu Leu Ala Ser Phe Arg Val Leu Pro Glu Ala Cys Glu Arg Leu65 70 75 80atc cgc agt ctt ccg gcc gaa tta gac gat atg gcg gtt ctg cgc acg 288Ile Arg Ser Leu Pro Ala Glu Leu Asp Asp Met Ala Val Leu Arg Thr 85 90 95gcc gtc tct tca ttc ggg gac ata tcg ttt gcg ttc aag ccg aca atg 336Ala Val Ser Ser Phe Gly Asp Ile Ser Phe Ala Phe Lys Pro Thr Met 100 105 110gag cag gcc ctc agg ctg att gcg gcc gtc ccg tca att atc gcg ttc 384Glu Gln Ala Leu Arg Leu Ile Ala Ala Val Pro Ser Ile Ile Ala Phe 115 120 125aga aag agg ctg gct gat gga gaa gag cct gtt cag ccg ctg gat gag 432Arg Lys Arg Leu Ala Asp Gly Glu Glu Pro Val Gln Pro Leu Asp Glu 130 135 140cta ggt ttt gtg gaa aat tat ttt tac atg ctg aaa gga aag cgt ccg 480Leu Gly Phe Val Glu Asn Tyr Phe Tyr Met Leu Lys Gly Lys Arg Pro145 150 155 160aca gaa gct cag aaa aaa gcg ctt gaa acg tac atg att ctt gcg atg 528Thr Glu Ala Gln Lys Lys Ala Leu Glu Thr Tyr Met Ile Leu Ala Met 165 170 175gaa cat ggc atg aac gct tca acc ttt tcg gca agg gtt acg gtg tct 576Glu His Gly Met Asn Ala Ser Thr Phe Ser Ala Arg Val Thr Val Ser 180 185 190acg gaa tcc gat ctt gtt tct gcc gtc aca tct gca ctt ggc acg atg 624Thr Glu Ser Asp Leu Val Ser Ala Val Thr Ser Ala Leu Gly Thr Met 195 200 205aaa gga cct ttg cac ggc gga gcg ccg tct gcg gtg acg gat atg ctg 672Lys Gly Pro Leu His Gly Gly Ala Pro Ser Ala Val Thr Asp Met Leu 210 215 220gaa ggc atc ggg gaa aaa cag cac gct gaa agc tat ata aga gaa aag 720Glu Gly Ile Gly Glu Lys Gln His Ala Glu Ser Tyr Ile Arg Glu Lys225 230 235 240ctt gaa aaa ggc gag aga ctg atg gga ttt gga cac cgc gtc tat aaa 768Leu Glu Lys Gly Glu Arg Leu Met Gly Phe Gly His Arg Val Tyr Lys 245 250 255acg cat gat ccg aga gcg aag gct tta agg gtg aaa gcc gag gaa att 816Thr His Asp Pro Arg Ala Lys Ala Leu Arg Val Lys Ala Glu Glu Ile 260 265 270gcc ggg ggc gac agg gat atc gac ctt gct ctt cat gtt gaa aat acg 864Ala Gly Gly Asp Arg Asp Ile Asp Leu Ala Leu His Val Glu Asn Thr 275 280 285gcg atc cgc ctt ttg gaa gaa tac aag ccg ggc cgc aag ctt tat acg 912Ala Ile Arg Leu Leu Glu Glu Tyr Lys Pro Gly Arg Lys Leu Tyr Thr 290 295 300aat gtc gaa ttt tac gca gcc gct gtg atg aag gcg atc gat ttt gat 960Asn Val Glu Phe Tyr Ala Ala Ala Val Met Lys Ala Ile Asp Phe Asp305 310 315 320gcc gca ttg ttc acc cct act ttt tcg gca gcg aga atg gtc ggc tgg 1008Ala Ala Leu Phe Thr Pro Thr Phe Ser Ala Ala Arg Met Val Gly Trp 325 330 335tgc gcc cat gtg ctt gag cag gct gac aac aac atg att ttc agg cca 1056Cys Ala His Val Leu Glu Gln Ala Asp Asn Asn Met Ile Phe Arg Pro 340 345 350tcg gct aaa tat gtc gga gaa ttg gtc tca cag taa 1092Ser Ala Lys Tyr Val Gly Glu Leu Val Ser Gln 355 36060363PRTBacillus licheniformis DSM 13 60Met Val Tyr His Gly Leu Lys Gly Ile Thr Cys Val Glu Thr Ala Ile1 5 10 15Ser Arg Ile Asp Gly Glu Lys Gly Lys Leu Ile Tyr Arg Gly Tyr Asn 20 25 30Ala Asp Glu Leu Ala Leu Ser Ser Thr Phe Glu Glu Val Ala Tyr Leu 35 40 45Ile Leu Asn Gly Ala Phe Pro Thr Glu Ala Glu Leu Ser Arg Phe Asn 50 55 60Glu Leu Leu Ala Ser Phe Arg Val Leu Pro Glu Ala Cys Glu Arg Leu65 70 75 80Ile Arg Ser Leu Pro Ala Glu Leu Asp Asp Met Ala Val Leu Arg Thr 85 90 95Ala Val Ser Ser Phe Gly Asp Ile Ser Phe Ala Phe Lys Pro Thr Met 100 105 110Glu Gln Ala Leu Arg Leu Ile Ala Ala Val Pro Ser Ile Ile Ala Phe 115 120 125Arg Lys Arg Leu Ala Asp Gly Glu Glu Pro Val Gln Pro Leu Asp Glu 130 135 140Leu Gly Phe Val Glu Asn Tyr Phe Tyr Met Leu Lys Gly Lys Arg Pro145 150 155 160Thr Glu Ala Gln Lys Lys Ala Leu Glu Thr Tyr Met Ile Leu Ala Met 165 170 175Glu His Gly Met Asn Ala Ser Thr Phe Ser Ala Arg Val Thr Val Ser 180 185 190Thr Glu Ser Asp Leu Val Ser Ala Val Thr Ser Ala Leu Gly Thr Met 195 200 205Lys Gly Pro Leu His Gly Gly Ala Pro Ser Ala Val Thr Asp Met Leu 210 215 220Glu Gly Ile Gly Glu Lys Gln His Ala Glu Ser Tyr Ile Arg Glu Lys225 230 235 240Leu Glu Lys Gly Glu Arg Leu Met Gly Phe Gly His Arg Val Tyr Lys 245 250 255Thr His Asp Pro Arg Ala Lys Ala Leu Arg Val Lys Ala Glu Glu Ile 260 265 270Ala Gly Gly Asp Arg Asp Ile Asp Leu Ala Leu His Val Glu Asn Thr 275 280 285Ala Ile Arg Leu Leu Glu Glu Tyr Lys Pro Gly Arg Lys Leu Tyr Thr 290 295 300Asn Val Glu Phe Tyr Ala Ala Ala Val Met Lys Ala Ile Asp Phe Asp305 310 315 320Ala Ala Leu Phe Thr Pro Thr Phe Ser Ala Ala Arg Met Val Gly Trp 325 330 335Cys Ala His Val Leu Glu Gln Ala Asp Asn Asn Met Ile Phe Arg Pro 340 345 350Ser Ala Lys Tyr Val Gly Glu Leu Val Ser Gln 355 36061879DNABacillus licheniformis DSM 13gene(1)..(879)alsR - transcriptional regulator of the alpha-acetolactate operon - BLi03849 61atg gag ctg cgc cat ctt cgt tat ttt atg gtt gtt gcc gaa gaa ctg 48Met Glu Leu Arg His Leu Arg Tyr Phe Met Val Val Ala Glu Glu Leu1 5 10 15cat ttc gga aaa gca gct ctt cgg ctg aac atg act cag cct ccg ctc 96His Phe Gly Lys Ala Ala Leu Arg Leu Asn Met Thr Gln Pro Pro Leu 20 25 30agc cag cag att aaa cag cta gaa agt gaa ata ggc gtc acc ttg ctg 144Ser Gln Gln Ile Lys Gln Leu Glu Ser Glu Ile Gly Val Thr Leu Leu 35 40 45aag agg tca aaa aga gcc gtc agg cta acg gcg gca ggc ggg gtc ttt 192Lys Arg Ser Lys Arg Ala Val Arg Leu Thr Ala Ala Gly Gly Val Phe 50 55 60tta aaa caa atc aag gaa gga ctt tcc cag att gat cag gcc gtc gac 240Leu Lys Gln Ile Lys Glu Gly Leu Ser Gln Ile Asp Gln Ala Val Asp65 70 75 80atg gct cag cgg acg gcc cgc ggg gag ctg gga aga ctc gtg atc ggc 288Met Ala Gln Arg Thr Ala Arg Gly Glu Leu Gly Arg Leu Val Ile Gly 85 90 95ttt gtc ggc tcc gca aca tac gag atc atg cct ccc atc atc agg gaa 336Phe Val Gly Ser Ala Thr Tyr Glu Ile Met Pro Pro Ile Ile Arg Glu 100 105 110tac cga aac cgg ttc ccg tcc gtg cag atc gat ctt cgg gag ctt tca 384Tyr Arg Asn Arg Phe Pro Ser Val Gln Ile Asp Leu Arg Glu Leu Ser 115 120 125acc ccc aat cag atc gat gca ctg cta aac ggg cat att gac atc ggg 432Thr Pro Asn Gln Ile Asp Ala Leu Leu Asn Gly His Ile Asp Ile Gly 130 135 140gtg ctg cat ccc ccg ttg gga aac gac gaa ctg aag tca tac acg gta 480Val Leu His Pro Pro Leu Gly Asn Asp Glu Leu Lys Ser Tyr Thr Val145 150 155 160aaa aaa agc cat tgt gtc ctt gca ctt cca aaa cac cac ccg ttg aca 528Lys Lys Ser His Cys Val Leu Ala Leu Pro Lys His His Pro Leu Thr 165 170 175gaa aaa gcc cgg gta cat ttg aag gat ctt gaa gaa gaa tcg ctg atc 576Glu Lys Ala Arg Val His Leu Lys Asp Leu Glu Glu Glu Ser Leu Ile 180 185 190gtc att gcc aaa gaa gcg tgg ccc tct tta tac acg gaa ttc aat ttc 624Val Ile Ala Lys Glu Ala Trp Pro Ser Leu Tyr Thr Glu Phe Asn Phe 195 200 205ttg tgt gaa aaa gcc gga ttt atc ccc aac atc gcc cag gaa gct aca 672Leu Cys Glu Lys Ala Gly Phe Ile Pro Asn Ile Ala Gln Glu Ala Thr 210 215 220gaa tac caa atg gtg atc ggc ctc gta tcg gcc gga atg gga atc gcc 720Glu Tyr Gln Met Val Ile Gly Leu Val Ser Ala Gly Met Gly Ile Ala225 230 235 240gtc gtg cct acg gcg gct aaa cga tta ttt aac ctt gat gtt gta tac 768Val Val Pro Thr Ala Ala Lys Arg Leu Phe Asn Leu Asp Val Val Tyr 245 250 255aaa gaa atc gaa gac ttt ccc ctt cgc gcc gaa tgg att acc gca cat 816Lys Glu Ile Glu Asp Phe Pro Leu Arg Ala Glu Trp Ile Thr Ala His 260 265 270cga aag gac aac cgc aat ccg gca ctt aag cat ttc atc gaa atc tca 864Arg Lys Asp Asn Arg Asn Pro Ala Leu Lys His Phe Ile Glu Ile Ser 275 280 285gat caa cac tca tag 879Asp Gln His Ser 29062292PRTBacillus licheniformis DSM 13 62Met Glu Leu Arg His Leu Arg Tyr Phe Met Val Val Ala Glu Glu Leu1 5 10 15His Phe Gly Lys Ala Ala Leu Arg Leu Asn Met Thr Gln Pro Pro Leu 20 25 30Ser Gln Gln Ile Lys Gln Leu Glu Ser Glu Ile Gly Val Thr Leu Leu 35 40 45Lys Arg Ser Lys Arg Ala Val Arg Leu Thr Ala Ala Gly Gly Val Phe 50 55 60Leu Lys Gln Ile Lys Glu Gly Leu Ser Gln Ile Asp Gln Ala Val Asp65 70 75 80Met Ala Gln Arg Thr Ala Arg Gly Glu Leu Gly Arg Leu Val Ile Gly 85 90 95Phe Val Gly Ser Ala Thr Tyr Glu Ile Met Pro Pro Ile Ile Arg Glu 100 105 110Tyr Arg Asn Arg Phe Pro Ser Val Gln Ile Asp Leu Arg Glu Leu Ser 115 120 125Thr Pro Asn Gln Ile Asp Ala Leu Leu Asn Gly His Ile Asp Ile Gly 130 135 140Val Leu His Pro Pro Leu Gly Asn Asp Glu Leu Lys Ser Tyr Thr Val145 150 155 160Lys Lys Ser His Cys Val Leu Ala Leu Pro Lys His His Pro Leu Thr 165 170 175Glu Lys Ala Arg Val His Leu Lys Asp Leu Glu Glu Glu Ser Leu Ile 180 185 190Val Ile Ala Lys Glu Ala Trp Pro Ser Leu Tyr Thr Glu Phe Asn Phe 195 200 205Leu Cys Glu Lys Ala Gly Phe Ile Pro Asn Ile Ala Gln Glu Ala Thr 210 215 220Glu Tyr Gln Met Val Ile Gly Leu Val Ser Ala Gly Met Gly Ile Ala225 230 235 240Val Val Pro Thr Ala Ala Lys Arg Leu Phe Asn Leu Asp Val Val Tyr 245 250 255Lys Glu Ile Glu Asp Phe Pro Leu Arg Ala Glu Trp Ile Thr Ala His 260 265 270Arg Lys Asp Asn Arg Asn Pro Ala Leu Lys His Phe Ile Glu Ile Ser 275 280 285Asp Gln His Ser 290631380DNABacillus licheniformis DSM

13gene(1)..(1380)putative protein - putative ammonium transporter - BLi01175 63atg aaa acc aag gta tct gcc gtc ctg ttc ctc agt ttt ttc ctc gca 48Met Lys Thr Lys Val Ser Ala Val Leu Phe Leu Ser Phe Phe Leu Ala1 5 10 15gca tca gct ctg gcc gcc gaa ccg acg gtg caa tcg gtc aat gac tcg 96Ala Ser Ala Leu Ala Ala Glu Pro Thr Val Gln Ser Val Asn Asp Ser 20 25 30gtc aac atg aca tgg gta gcc atc ggc act ctg ctc gtc ttt ttt atg 144Val Asn Met Thr Trp Val Ala Ile Gly Thr Leu Leu Val Phe Phe Met 35 40 45cac gcc ggt ttt gcg atg gtt gaa tca ggc ttt aca aga gct aaa aac 192His Ala Gly Phe Ala Met Val Glu Ser Gly Phe Thr Arg Ala Lys Asn 50 55 60acg tta aac att tta atg aaa aac ttc ctg acg atc tct atc ggc tcc 240Thr Leu Asn Ile Leu Met Lys Asn Phe Leu Thr Ile Ser Ile Gly Ser65 70 75 80atc ctt tat ttc ttc gcc gga tac gcg ttc atg ttc ggg gac agc gcc 288Ile Leu Tyr Phe Phe Ala Gly Tyr Ala Phe Met Phe Gly Asp Ser Ala 85 90 95ggc ggg ttc atc ggc aca agc ggt ttt gct cta gcg ggt gat cag gat 336Gly Gly Phe Ile Gly Thr Ser Gly Phe Ala Leu Ala Gly Asp Gln Asp 100 105 110gtc gga ttt ttc gtc ttt cag gcc gtg ttt gcc gct acg tgc gcc acc 384Val Gly Phe Phe Val Phe Gln Ala Val Phe Ala Ala Thr Cys Ala Thr 115 120 125att att tcc ggc gct gtc gct gaa cgg atg aag ctt ggc agc tac atg 432Ile Ile Ser Gly Ala Val Ala Glu Arg Met Lys Leu Gly Ser Tyr Met 130 135 140atc ttg acg ctg ttt atg acc ggt ctc atc tat ccg gtt gtc ggc cat 480Ile Leu Thr Leu Phe Met Thr Gly Leu Ile Tyr Pro Val Val Gly His145 150 155 160tgg act tgg ggg gaa ggc tgg ctg tac gac ctc gga ttt gtc gat ttt 528Trp Thr Trp Gly Glu Gly Trp Leu Tyr Asp Leu Gly Phe Val Asp Phe 165 170 175gcc gga tct tca atc gtt cac ttg aca ggc gct gcg ggc gca gtt gcg 576Ala Gly Ser Ser Ile Val His Leu Thr Gly Ala Ala Gly Ala Val Ala 180 185 190gct gtc atg ttt ctc ggg ccg cgg atc gga aaa tat aca aac ggc aag 624Ala Val Met Phe Leu Gly Pro Arg Ile Gly Lys Tyr Thr Asn Gly Lys 195 200 205gtc aac gcc att cca ggg cac agc att ccg ctg ggc gct ctc ggc gtc 672Val Asn Ala Ile Pro Gly His Ser Ile Pro Leu Gly Ala Leu Gly Val 210 215 220ttt atc ctg tgg ttc ggc tgg ttt ggg ttc aac ggc gga agc acg ctt 720Phe Ile Leu Trp Phe Gly Trp Phe Gly Phe Asn Gly Gly Ser Thr Leu225 230 235 240gcg gct gat ccc gcc gcc gtc ccg cac gtc atc acc aca acc ctt tta 768Ala Ala Asp Pro Ala Ala Val Pro His Val Ile Thr Thr Thr Leu Leu 245 250 255gcg gca tcg ggc gga gtc gtc agc tcg gcg ctg ttc act tat ttc cgc 816Ala Ala Ser Gly Gly Val Val Ser Ser Ala Leu Phe Thr Tyr Phe Arg 260 265 270tac ggc agg atc gat cct tcc ctg acg ctg aac ggc gcg cta ggc ggc 864Tyr Gly Arg Ile Asp Pro Ser Leu Thr Leu Asn Gly Ala Leu Gly Gly 275 280 285ctt gtc gga att acc gcg ggc gcg gac ggc gtt tca ttt aca ggc tcg 912Leu Val Gly Ile Thr Ala Gly Ala Asp Gly Val Ser Phe Thr Gly Ser 290 295 300ata ttg atc gga ttc atc gcc ggt gtc att tta gtt gtc ggc gtt cag 960Ile Leu Ile Gly Phe Ile Ala Gly Val Ile Leu Val Val Gly Val Gln305 310 315 320ttt atc gac cag gtg ctg aag ctt gac gat ccc gtc ggc gcc ctt gcc 1008Phe Ile Asp Gln Val Leu Lys Leu Asp Asp Pro Val Gly Ala Leu Ala 325 330 335gtt cac ggt att tgc ggt gtt tgg ggg acg ctt gca gtc ggg ctt ttc 1056Val His Gly Ile Cys Gly Val Trp Gly Thr Leu Ala Val Gly Leu Phe 340 345 350aat aca tcg agc ggt ctc ttt tac gga gga gga gcc gca cag ctt ggc 1104Asn Thr Ser Ser Gly Leu Phe Tyr Gly Gly Gly Ala Ala Gln Leu Gly 355 360 365att caa gcc gtc ggc att ttg tcg atc gcg gcc tgg acc ctc gct gcc 1152Ile Gln Ala Val Gly Ile Leu Ser Ile Ala Ala Trp Thr Leu Ala Ala 370 375 380aca tac gct gtc gcc gcg gca atg aaa gcc ctc gct ccg atc agg gta 1200Thr Tyr Ala Val Ala Ala Ala Met Lys Ala Leu Ala Pro Ile Arg Val385 390 395 400tca aaa gag gag gaa att tcc gga cta gat tat gcc gaa cac ggt tct 1248Ser Lys Glu Glu Glu Ile Ser Gly Leu Asp Tyr Ala Glu His Gly Ser 405 410 415tcc gcg tac gag tat aaa gaa agc tac atc gat cag gac gat gtt gac 1296Ser Ala Tyr Glu Tyr Lys Glu Ser Tyr Ile Asp Gln Asp Asp Val Asp 420 425 430gta agc tcc gct ttc gga acg gat ctt gtc tca cgg ctc aac aat tta 1344Val Ser Ser Ala Phe Gly Thr Asp Leu Val Ser Arg Leu Asn Asn Leu 435 440 445aag cct gga cct tct tcc aaa act gaa cag gtg tga 1380Lys Pro Gly Pro Ser Ser Lys Thr Glu Gln Val 450 45564459PRTBacillus licheniformis DSM 13 64Met Lys Thr Lys Val Ser Ala Val Leu Phe Leu Ser Phe Phe Leu Ala1 5 10 15Ala Ser Ala Leu Ala Ala Glu Pro Thr Val Gln Ser Val Asn Asp Ser 20 25 30Val Asn Met Thr Trp Val Ala Ile Gly Thr Leu Leu Val Phe Phe Met 35 40 45His Ala Gly Phe Ala Met Val Glu Ser Gly Phe Thr Arg Ala Lys Asn 50 55 60Thr Leu Asn Ile Leu Met Lys Asn Phe Leu Thr Ile Ser Ile Gly Ser65 70 75 80Ile Leu Tyr Phe Phe Ala Gly Tyr Ala Phe Met Phe Gly Asp Ser Ala 85 90 95Gly Gly Phe Ile Gly Thr Ser Gly Phe Ala Leu Ala Gly Asp Gln Asp 100 105 110Val Gly Phe Phe Val Phe Gln Ala Val Phe Ala Ala Thr Cys Ala Thr 115 120 125Ile Ile Ser Gly Ala Val Ala Glu Arg Met Lys Leu Gly Ser Tyr Met 130 135 140Ile Leu Thr Leu Phe Met Thr Gly Leu Ile Tyr Pro Val Val Gly His145 150 155 160Trp Thr Trp Gly Glu Gly Trp Leu Tyr Asp Leu Gly Phe Val Asp Phe 165 170 175Ala Gly Ser Ser Ile Val His Leu Thr Gly Ala Ala Gly Ala Val Ala 180 185 190Ala Val Met Phe Leu Gly Pro Arg Ile Gly Lys Tyr Thr Asn Gly Lys 195 200 205Val Asn Ala Ile Pro Gly His Ser Ile Pro Leu Gly Ala Leu Gly Val 210 215 220Phe Ile Leu Trp Phe Gly Trp Phe Gly Phe Asn Gly Gly Ser Thr Leu225 230 235 240Ala Ala Asp Pro Ala Ala Val Pro His Val Ile Thr Thr Thr Leu Leu 245 250 255Ala Ala Ser Gly Gly Val Val Ser Ser Ala Leu Phe Thr Tyr Phe Arg 260 265 270Tyr Gly Arg Ile Asp Pro Ser Leu Thr Leu Asn Gly Ala Leu Gly Gly 275 280 285Leu Val Gly Ile Thr Ala Gly Ala Asp Gly Val Ser Phe Thr Gly Ser 290 295 300Ile Leu Ile Gly Phe Ile Ala Gly Val Ile Leu Val Val Gly Val Gln305 310 315 320Phe Ile Asp Gln Val Leu Lys Leu Asp Asp Pro Val Gly Ala Leu Ala 325 330 335Val His Gly Ile Cys Gly Val Trp Gly Thr Leu Ala Val Gly Leu Phe 340 345 350Asn Thr Ser Ser Gly Leu Phe Tyr Gly Gly Gly Ala Ala Gln Leu Gly 355 360 365Ile Gln Ala Val Gly Ile Leu Ser Ile Ala Ala Trp Thr Leu Ala Ala 370 375 380Thr Tyr Ala Val Ala Ala Ala Met Lys Ala Leu Ala Pro Ile Arg Val385 390 395 400Ser Lys Glu Glu Glu Ile Ser Gly Leu Asp Tyr Ala Glu His Gly Ser 405 410 415Ser Ala Tyr Glu Tyr Lys Glu Ser Tyr Ile Asp Gln Asp Asp Val Asp 420 425 430Val Ser Ser Ala Phe Gly Thr Asp Leu Val Ser Arg Leu Asn Asn Leu 435 440 445Lys Pro Gly Pro Ser Ser Lys Thr Glu Gln Val 450 455651032DNABacillus licheniformis DSM 13gene(1)..(1032)ycdH - unknown function - similar to ABC transporter (binding protein) - BLi03213 65atg aaa aaa aca ttc gga ata gca tct gca ttc att ctt gct gcc gga 48Met Lys Lys Thr Phe Gly Ile Ala Ser Ala Phe Ile Leu Ala Ala Gly1 5 10 15tta aca gcg ggt tgc tca agc aat gga gct tcc agt gaa caa aaa gaa 96Leu Thr Ala Gly Cys Ser Ser Asn Gly Ala Ser Ser Glu Gln Lys Glu 20 25 30aat aaa gac acg ctg gac atc tat aca acc att tat ccg ctt gaa gac 144Asn Lys Asp Thr Leu Asp Ile Tyr Thr Thr Ile Tyr Pro Leu Glu Asp 35 40 45ttc acc aaa aag atc ggc gga gaa tat gtg aac gtt aaa agt gtt tac 192Phe Thr Lys Lys Ile Gly Gly Glu Tyr Val Asn Val Lys Ser Val Tyr 50 55 60ccg cca aac gtt gaa gcg cac act tac gag cca agc tca aaa acg atg 240Pro Pro Asn Val Glu Ala His Thr Tyr Glu Pro Ser Ser Lys Thr Met65 70 75 80gcg gaa att tcc gac tcg gat gcc ttt ata tac tct gga gtc ggt gtt 288Ala Glu Ile Ser Asp Ser Asp Ala Phe Ile Tyr Ser Gly Val Gly Val 85 90 95gaa gga ttc gca gac aaa gct gta agc aca ctt aaa aac agc gac gtc 336Glu Gly Phe Ala Asp Lys Ala Val Ser Thr Leu Lys Asn Ser Asp Val 100 105 110aaa atc gtc aaa gcc ggg gag ggc att gac ctt ctg tca cgt gaa gaa 384Lys Ile Val Lys Ala Gly Glu Gly Ile Asp Leu Leu Ser Arg Glu Glu 115 120 125gaa ggc gaa cat gat gag cat gca cat gga gaa gaa cac ggt gag cac 432Glu Gly Glu His Asp Glu His Ala His Gly Glu Glu His Gly Glu His 130 135 140gat gag cat gca cat gaa gaa gaa cat gaa cac gga gaa gcc gaa ggt 480Asp Glu His Ala His Glu Glu Glu His Glu His Gly Glu Ala Glu Gly145 150 155 160cac gat cat cat cac gac cat gga gat cat gac ccg cac gca tgg ctt 528His Asp His His His Asp His Gly Asp His Asp Pro His Ala Trp Leu 165 170 175gat cct gtc ctt gct gaa aaa atg gcc gaa aac att aaa gac caa ctc 576Asp Pro Val Leu Ala Glu Lys Met Ala Glu Asn Ile Lys Asp Gln Leu 180 185 190gta aaa tta gaa cct gaa cac gaa aaa acg ttt acg aaa aac tat gag 624Val Lys Leu Glu Pro Glu His Glu Lys Thr Phe Thr Lys Asn Tyr Glu 195 200 205tca tta aaa aaa gac ttg gag cag ctt gac aaa gac ttc aaa caa aca 672Ser Leu Lys Lys Asp Leu Glu Gln Leu Asp Lys Asp Phe Lys Gln Thr 210 215 220gtc gac aaa gct gat aaa aaa gaa att ctc gtc tca cat gcc gcc tac 720Val Asp Lys Ala Asp Lys Lys Glu Ile Leu Val Ser His Ala Ala Tyr225 230 235 240ggc tac tgg gaa aaa cgc tac ggc att aag cag ata agc gtt ctc ggc 768Gly Tyr Trp Glu Lys Arg Tyr Gly Ile Lys Gln Ile Ser Val Leu Gly 245 250 255ctt tcg cct act gag gaa cct tca caa aag cag ctt gaa aac att gtt 816Leu Ser Pro Thr Glu Glu Pro Ser Gln Lys Gln Leu Glu Asn Ile Val 260 265 270aaa aca gcg aaa gaa cat aat att aaa tat atc att ttt gaa agc aat 864Lys Thr Ala Lys Glu His Asn Ile Lys Tyr Ile Ile Phe Glu Ser Asn 275 280 285gtc agc agc aaa atc tcc gag atc atc aaa aaa gaa atc gga gcg gaa 912Val Ser Ser Lys Ile Ser Glu Ile Ile Lys Lys Glu Ile Gly Ala Glu 290 295 300agt ctg acg atg aaa aac ctt gaa tct gta aca aaa gaa gac att gaa 960Ser Leu Thr Met Lys Asn Leu Glu Ser Val Thr Lys Glu Asp Ile Glu305 310 315 320agc ggc aaa gat tat acc gcc atc atg aaa gaa aac ctg gac gtc ttg 1008Ser Gly Lys Asp Tyr Thr Ala Ile Met Lys Glu Asn Leu Asp Val Leu 325 330 335aaa aaa gct tta tca agc aaa taa 1032Lys Lys Ala Leu Ser Ser Lys 34066343PRTBacillus licheniformis DSM 13 66Met Lys Lys Thr Phe Gly Ile Ala Ser Ala Phe Ile Leu Ala Ala Gly1 5 10 15Leu Thr Ala Gly Cys Ser Ser Asn Gly Ala Ser Ser Glu Gln Lys Glu 20 25 30Asn Lys Asp Thr Leu Asp Ile Tyr Thr Thr Ile Tyr Pro Leu Glu Asp 35 40 45Phe Thr Lys Lys Ile Gly Gly Glu Tyr Val Asn Val Lys Ser Val Tyr 50 55 60Pro Pro Asn Val Glu Ala His Thr Tyr Glu Pro Ser Ser Lys Thr Met65 70 75 80Ala Glu Ile Ser Asp Ser Asp Ala Phe Ile Tyr Ser Gly Val Gly Val 85 90 95Glu Gly Phe Ala Asp Lys Ala Val Ser Thr Leu Lys Asn Ser Asp Val 100 105 110Lys Ile Val Lys Ala Gly Glu Gly Ile Asp Leu Leu Ser Arg Glu Glu 115 120 125Glu Gly Glu His Asp Glu His Ala His Gly Glu Glu His Gly Glu His 130 135 140Asp Glu His Ala His Glu Glu Glu His Glu His Gly Glu Ala Glu Gly145 150 155 160His Asp His His His Asp His Gly Asp His Asp Pro His Ala Trp Leu 165 170 175Asp Pro Val Leu Ala Glu Lys Met Ala Glu Asn Ile Lys Asp Gln Leu 180 185 190Val Lys Leu Glu Pro Glu His Glu Lys Thr Phe Thr Lys Asn Tyr Glu 195 200 205Ser Leu Lys Lys Asp Leu Glu Gln Leu Asp Lys Asp Phe Lys Gln Thr 210 215 220Val Asp Lys Ala Asp Lys Lys Glu Ile Leu Val Ser His Ala Ala Tyr225 230 235 240Gly Tyr Trp Glu Lys Arg Tyr Gly Ile Lys Gln Ile Ser Val Leu Gly 245 250 255Leu Ser Pro Thr Glu Glu Pro Ser Gln Lys Gln Leu Glu Asn Ile Val 260 265 270Lys Thr Ala Lys Glu His Asn Ile Lys Tyr Ile Ile Phe Glu Ser Asn 275 280 285Val Ser Ser Lys Ile Ser Glu Ile Ile Lys Lys Glu Ile Gly Ala Glu 290 295 300Ser Leu Thr Met Lys Asn Leu Glu Ser Val Thr Lys Glu Asp Ile Glu305 310 315 320Ser Gly Lys Asp Tyr Thr Ala Ile Met Lys Glu Asn Leu Asp Val Leu 325 330 335Lys Lys Ala Leu Ser Ser Lys 340671548DNABacillus licheniformis DSM 13gene(1)..(1548)trpE - anthranilate synthase - BLi02403 67ttg agt ctg caa acc gat ttt gcc tct ttt tca gag gat tct tca cac 48Leu Ser Leu Gln Thr Asp Phe Ala Ser Phe Ser Glu Asp Ser Ser His1 5 10 15ttt aaa acg ata ccg att att gag agt ttt tcc gtc gat acg ctg acg 96Phe Lys Thr Ile Pro Ile Ile Glu Ser Phe Ser Val Asp Thr Leu Thr 20 25 30ccc gtt caa ttg gtc gag aag ctg cag gaa gac atc gtc tat ttg ctg 144Pro Val Gln Leu Val Glu Lys Leu Gln Glu Asp Ile Val Tyr Leu Leu 35 40 45gaa agc cag gat gaa acc tct gca tgg tca aga tat tct ttc atc ggc 192Glu Ser Gln Asp Glu Thr Ser Ala Trp Ser Arg Tyr Ser Phe Ile Gly 50 55 60ctt gac ccc ttt ttg acg atc aaa gaa gaa cgc ggc cgg ttt cac gcc 240Leu Asp Pro Phe Leu Thr Ile Lys Glu Glu Arg Gly Arg Phe His Ala65 70 75 80tgc gat caa aag cag aag aag ctt tat tcg gct tca cag tta aaa gac 288Cys Asp Gln Lys Gln Lys Lys Leu Tyr Ser Ala Ser Gln Leu Lys Asp 85 90 95gtt ctc gaa tgg atg tcg gag acg tat tcg atc aaa gcg cct gag ctc 336Val Leu Glu Trp Met Ser Glu Thr Tyr Ser Ile Lys Ala Pro Glu Leu 100 105 110gac atc ccg ttt aca gga gga gcg gtc ggc tat tta agc tac gat tta 384Asp Ile Pro Phe Thr Gly Gly Ala Val Gly Tyr Leu Ser Tyr Asp Leu 115 120 125atg ccg ctc att gaa ccg tcc atc aaa ggg cac agc aaa gaa acg gaa 432Met Pro Leu Ile Glu Pro Ser Ile Lys Gly His Ser Lys Glu Thr Glu 130 135 140atg gca aaa tgc ctc ctg ttt gtc tgc cgg acg atg atc gct tat gat 480Met Ala Lys Cys Leu Leu Phe Val Cys Arg Thr Met Ile Ala Tyr Asp145 150 155 160cac gaa ctg aaa aag ctg cat ttc atc cac tat aca agg ctt gaa ggc 528His Glu Leu Lys Lys Leu His Phe Ile His Tyr Thr Arg Leu Glu Gly 165 170 175agt gaa aat agg gag gaa aag cgc cgc att ttc aaa aag ggg aaa gaa 576Ser Glu Asn Arg Glu Glu Lys Arg Arg Ile Phe Lys Lys Gly Lys Glu 180 185 190gag ctc aag cag ctt aca cac agg ctg tca gac cgg aaa acg ccg aaa 624Glu Leu Lys Gln Leu Thr His Arg Leu Ser Asp Arg Lys Thr Pro Lys

195 200 205gag ctg ttt ctt ccg caa gga cgg cac acg gcc ccc agc ttt gaa ggg 672Glu Leu Phe Leu Pro Gln Gly Arg His Thr Ala Pro Ser Phe Glu Gly 210 215 220att caa tca aca tat gag aaa tcc cgg ttt ttg gcc gat gtc gat cgc 720Ile Gln Ser Thr Tyr Glu Lys Ser Arg Phe Leu Ala Asp Val Asp Arg225 230 235 240ttg aaa gaa tac att aaa gcg gga gac att ttt caa ggc gtc ctt tcg 768Leu Lys Glu Tyr Ile Lys Ala Gly Asp Ile Phe Gln Gly Val Leu Ser 245 250 255caa agg ttc gac att ccg gtg aag gtc ggt tct ttt gag tta tac aga 816Gln Arg Phe Asp Ile Pro Val Lys Val Gly Ser Phe Glu Leu Tyr Arg 260 265 270gtg ctg aga atg atc aac ccg tcc ccg tac atg tac tac atg agg ctt 864Val Leu Arg Met Ile Asn Pro Ser Pro Tyr Met Tyr Tyr Met Arg Leu 275 280 285cct gac cgc gat ctc gtc gga agc tcg ccc gaa cgt ctg atc cat gtt 912Pro Asp Arg Asp Leu Val Gly Ser Ser Pro Glu Arg Leu Ile His Val 290 295 300caa aac ggc cat ctt gaa atc cat ccg atc gcc ggc aca agg aaa agg 960Gln Asn Gly His Leu Glu Ile His Pro Ile Ala Gly Thr Arg Lys Arg305 310 315 320ggc ggt gat cag gct gag gat aag cgg ctt gaa aaa gaa cta ttg gct 1008Gly Gly Asp Gln Ala Glu Asp Lys Arg Leu Glu Lys Glu Leu Leu Ala 325 330 335gat gag aaa gaa aaa gcg gag cat tac atg ctt gtc gat ttg gcg aga 1056Asp Glu Lys Glu Lys Ala Glu His Tyr Met Leu Val Asp Leu Ala Arg 340 345 350aat gat atc ggc aga gta gct gag tac ggg tct gtc caa gtg ccc gag 1104Asn Asp Ile Gly Arg Val Ala Glu Tyr Gly Ser Val Gln Val Pro Glu 355 360 365ttt acg aaa atc gtt tcc ttt tcg cac gtc atg cac atc att tcg atc 1152Phe Thr Lys Ile Val Ser Phe Ser His Val Met His Ile Ile Ser Ile 370 375 380gta acc ggc act tta aaa gcg ggt gtc cac ccg gtt gac gct tta atg 1200Val Thr Gly Thr Leu Lys Ala Gly Val His Pro Val Asp Ala Leu Met385 390 395 400agc gca ttt ccg gcc gga act ttg acc ggc gct ccg aaa atc cgc gcg 1248Ser Ala Phe Pro Ala Gly Thr Leu Thr Gly Ala Pro Lys Ile Arg Ala 405 410 415att cag ctt tta aat gag ctt gag ccc gaa ccg agg gaa aca tac gga 1296Ile Gln Leu Leu Asn Glu Leu Glu Pro Glu Pro Arg Glu Thr Tyr Gly 420 425 430gga tgc att gct tac atc gga ttt gac gga aat atc gat tcg tgc atc 1344Gly Cys Ile Ala Tyr Ile Gly Phe Asp Gly Asn Ile Asp Ser Cys Ile 435 440 445acg atc cgg acg atg agc gtg aaa aac gga gtg gcg tcc att cag gcc 1392Thr Ile Arg Thr Met Ser Val Lys Asn Gly Val Ala Ser Ile Gln Ala 450 455 460ggg gcg ggc atc gta gcc gac tcc gtg cct gaa aaa gaa tgg gaa gag 1440Gly Ala Gly Ile Val Ala Asp Ser Val Pro Glu Lys Glu Trp Glu Glu465 470 475 480acc tgc aat aaa gca ggg gct ctg tta aaa acg att caa atc gca gaa 1488Thr Cys Asn Lys Ala Gly Ala Leu Leu Lys Thr Ile Gln Ile Ala Glu 485 490 495gaa acg ttt ctc gaa aag ggg gag gac ggc aat gaa cga act gct gaa 1536Glu Thr Phe Leu Glu Lys Gly Glu Asp Gly Asn Glu Arg Thr Ala Glu 500 505 510atc atg cgt taa 1548Ile Met Arg 51568515PRTBacillus licheniformis DSM 13misc_feature(1)..(1)Xaa can be Leu or Met. 68Xaa Ser Leu Gln Thr Asp Phe Ala Ser Phe Ser Glu Asp Ser Ser His1 5 10 15Phe Lys Thr Ile Pro Ile Ile Glu Ser Phe Ser Val Asp Thr Leu Thr 20 25 30Pro Val Gln Leu Val Glu Lys Leu Gln Glu Asp Ile Val Tyr Leu Leu 35 40 45Glu Ser Gln Asp Glu Thr Ser Ala Trp Ser Arg Tyr Ser Phe Ile Gly 50 55 60Leu Asp Pro Phe Leu Thr Ile Lys Glu Glu Arg Gly Arg Phe His Ala65 70 75 80Cys Asp Gln Lys Gln Lys Lys Leu Tyr Ser Ala Ser Gln Leu Lys Asp 85 90 95Val Leu Glu Trp Met Ser Glu Thr Tyr Ser Ile Lys Ala Pro Glu Leu 100 105 110Asp Ile Pro Phe Thr Gly Gly Ala Val Gly Tyr Leu Ser Tyr Asp Leu 115 120 125Met Pro Leu Ile Glu Pro Ser Ile Lys Gly His Ser Lys Glu Thr Glu 130 135 140Met Ala Lys Cys Leu Leu Phe Val Cys Arg Thr Met Ile Ala Tyr Asp145 150 155 160His Glu Leu Lys Lys Leu His Phe Ile His Tyr Thr Arg Leu Glu Gly 165 170 175Ser Glu Asn Arg Glu Glu Lys Arg Arg Ile Phe Lys Lys Gly Lys Glu 180 185 190Glu Leu Lys Gln Leu Thr His Arg Leu Ser Asp Arg Lys Thr Pro Lys 195 200 205Glu Leu Phe Leu Pro Gln Gly Arg His Thr Ala Pro Ser Phe Glu Gly 210 215 220Ile Gln Ser Thr Tyr Glu Lys Ser Arg Phe Leu Ala Asp Val Asp Arg225 230 235 240Leu Lys Glu Tyr Ile Lys Ala Gly Asp Ile Phe Gln Gly Val Leu Ser 245 250 255Gln Arg Phe Asp Ile Pro Val Lys Val Gly Ser Phe Glu Leu Tyr Arg 260 265 270Val Leu Arg Met Ile Asn Pro Ser Pro Tyr Met Tyr Tyr Met Arg Leu 275 280 285Pro Asp Arg Asp Leu Val Gly Ser Ser Pro Glu Arg Leu Ile His Val 290 295 300Gln Asn Gly His Leu Glu Ile His Pro Ile Ala Gly Thr Arg Lys Arg305 310 315 320Gly Gly Asp Gln Ala Glu Asp Lys Arg Leu Glu Lys Glu Leu Leu Ala 325 330 335Asp Glu Lys Glu Lys Ala Glu His Tyr Met Leu Val Asp Leu Ala Arg 340 345 350Asn Asp Ile Gly Arg Val Ala Glu Tyr Gly Ser Val Gln Val Pro Glu 355 360 365Phe Thr Lys Ile Val Ser Phe Ser His Val Met His Ile Ile Ser Ile 370 375 380Val Thr Gly Thr Leu Lys Ala Gly Val His Pro Val Asp Ala Leu Met385 390 395 400Ser Ala Phe Pro Ala Gly Thr Leu Thr Gly Ala Pro Lys Ile Arg Ala 405 410 415Ile Gln Leu Leu Asn Glu Leu Glu Pro Glu Pro Arg Glu Thr Tyr Gly 420 425 430Gly Cys Ile Ala Tyr Ile Gly Phe Asp Gly Asn Ile Asp Ser Cys Ile 435 440 445Thr Ile Arg Thr Met Ser Val Lys Asn Gly Val Ala Ser Ile Gln Ala 450 455 460Gly Ala Gly Ile Val Ala Asp Ser Val Pro Glu Lys Glu Trp Glu Glu465 470 475 480Thr Cys Asn Lys Ala Gly Ala Leu Leu Lys Thr Ile Gln Ile Ala Glu 485 490 495Glu Thr Phe Leu Glu Lys Gly Glu Asp Gly Asn Glu Arg Thr Ala Glu 500 505 510Ile Met Arg 515691017DNABacillus licheniformis DSM 13gene(1)..(1017)trpD - anthranilate phosphoribosyltransferase - BLi02402 69atg aac gaa ctg ctg aaa tca tgc gtt aac gga cgg aca ctg aat gaa 48Met Asn Glu Leu Leu Lys Ser Cys Val Asn Gly Arg Thr Leu Asn Glu1 5 10 15atc gag gca cac agc atc atg cgc gac atg atg acc ggc tcg ctg agc 96Ile Glu Ala His Ser Ile Met Arg Asp Met Met Thr Gly Ser Leu Ser 20 25 30gct gcg gaa atc gga ggc ctc ctg tcc gtt ctc gcc tac agg gga gaa 144Ala Ala Glu Ile Gly Gly Leu Leu Ser Val Leu Ala Tyr Arg Gly Glu 35 40 45acg gct gaa gag att acg gga ttt gtc aga gcc atg cgg gag cag gcg 192Thr Ala Glu Glu Ile Thr Gly Phe Val Arg Ala Met Arg Glu Gln Ala 50 55 60cat aca ata gac gga ccg gaa cat gtt gtt gat acg tgc gga acc gga 240His Thr Ile Asp Gly Pro Glu His Val Val Asp Thr Cys Gly Thr Gly65 70 75 80gga gac gga agc tcc acc ttc aac att tcc aca gcg gct gcg atc atc 288Gly Asp Gly Ser Ser Thr Phe Asn Ile Ser Thr Ala Ala Ala Ile Ile 85 90 95gct tca tca gcc ggg gca aaa atc gca aaa cac ggt aat cga tcc gtt 336Ala Ser Ser Ala Gly Ala Lys Ile Ala Lys His Gly Asn Arg Ser Val 100 105 110tct tca aaa agc ggg agc gct gac gtt ctc gaa tat ctc ggc gtt tct 384Ser Ser Lys Ser Gly Ser Ala Asp Val Leu Glu Tyr Leu Gly Val Ser 115 120 125att cag acg tcg cct gaa gaa acg att aag agc att gaa cgc cga aat 432Ile Gln Thr Ser Pro Glu Glu Thr Ile Lys Ser Ile Glu Arg Arg Asn 130 135 140atg gga ttc ctg tat gcg cca agc tat cac tct tcc atg aag cac gtg 480Met Gly Phe Leu Tyr Ala Pro Ser Tyr His Ser Ser Met Lys His Val145 150 155 160gca ggc gcg agg aag gat ctg gct ttt aga acc gta ttc aat ctg ctc 528Ala Gly Ala Arg Lys Asp Leu Ala Phe Arg Thr Val Phe Asn Leu Leu 165 170 175ggc ccg ctc agc aac ccg atg aag gcc aaa cgc cag gtg atc ggc gtc 576Gly Pro Leu Ser Asn Pro Met Lys Ala Lys Arg Gln Val Ile Gly Val 180 185 190tac tcg atc gaa aaa gca aag ctt atg gcc cgc gct ctt gaa gcg ttt 624Tyr Ser Ile Glu Lys Ala Lys Leu Met Ala Arg Ala Leu Glu Ala Phe 195 200 205gaa ccg gag cac gta ttg ttt gtt tcg agc cgg gac ggt ctt gat gaa 672Glu Pro Glu His Val Leu Phe Val Ser Ser Arg Asp Gly Leu Asp Glu 210 215 220ctg tcg atc aca tcg ccg aca gat gta att gaa ctg aaa ggc ggc aaa 720Leu Ser Ile Thr Ser Pro Thr Asp Val Ile Glu Leu Lys Gly Gly Lys225 230 235 240aca tac gaa tac acc gtc gag ccg ggc gat gtt gga ctg cct ctc gga 768Thr Tyr Glu Tyr Thr Val Glu Pro Gly Asp Val Gly Leu Pro Leu Gly 245 250 255agg ctg aga gac att cag gtg gcg acg cct gaa gaa agc ggc cgt ctg 816Arg Leu Arg Asp Ile Gln Val Ala Thr Pro Glu Glu Ser Gly Arg Leu 260 265 270atc gtc gat att ttg aaa aac cgc ggt ccg gac tcg gct gtt cat atc 864Ile Val Asp Ile Leu Lys Asn Arg Gly Pro Asp Ser Ala Val His Ile 275 280 285gca gcg ttt aat gcc gga gcc gct tta tac gtc gca gaa cag gca gcg 912Ala Ala Phe Asn Ala Gly Ala Ala Leu Tyr Val Ala Glu Gln Ala Ala 290 295 300gat tta aaa gag ggc gtt gcg ctc gct tta gaa gct gtt gct ggc ggt 960Asp Leu Lys Glu Gly Val Ala Leu Ala Leu Glu Ala Val Ala Gly Gly305 310 315 320gaa gcg ctc aat cag ctg gag cga tta aaa cag aaa gag gaa gag ttg 1008Glu Ala Leu Asn Gln Leu Glu Arg Leu Lys Gln Lys Glu Glu Glu Leu 325 330 335tat gct taa 1017Tyr Ala70338PRTBacillus licheniformis DSM 13 70Met Asn Glu Leu Leu Lys Ser Cys Val Asn Gly Arg Thr Leu Asn Glu1 5 10 15Ile Glu Ala His Ser Ile Met Arg Asp Met Met Thr Gly Ser Leu Ser 20 25 30Ala Ala Glu Ile Gly Gly Leu Leu Ser Val Leu Ala Tyr Arg Gly Glu 35 40 45Thr Ala Glu Glu Ile Thr Gly Phe Val Arg Ala Met Arg Glu Gln Ala 50 55 60His Thr Ile Asp Gly Pro Glu His Val Val Asp Thr Cys Gly Thr Gly65 70 75 80Gly Asp Gly Ser Ser Thr Phe Asn Ile Ser Thr Ala Ala Ala Ile Ile 85 90 95Ala Ser Ser Ala Gly Ala Lys Ile Ala Lys His Gly Asn Arg Ser Val 100 105 110Ser Ser Lys Ser Gly Ser Ala Asp Val Leu Glu Tyr Leu Gly Val Ser 115 120 125Ile Gln Thr Ser Pro Glu Glu Thr Ile Lys Ser Ile Glu Arg Arg Asn 130 135 140Met Gly Phe Leu Tyr Ala Pro Ser Tyr His Ser Ser Met Lys His Val145 150 155 160Ala Gly Ala Arg Lys Asp Leu Ala Phe Arg Thr Val Phe Asn Leu Leu 165 170 175Gly Pro Leu Ser Asn Pro Met Lys Ala Lys Arg Gln Val Ile Gly Val 180 185 190Tyr Ser Ile Glu Lys Ala Lys Leu Met Ala Arg Ala Leu Glu Ala Phe 195 200 205Glu Pro Glu His Val Leu Phe Val Ser Ser Arg Asp Gly Leu Asp Glu 210 215 220Leu Ser Ile Thr Ser Pro Thr Asp Val Ile Glu Leu Lys Gly Gly Lys225 230 235 240Thr Tyr Glu Tyr Thr Val Glu Pro Gly Asp Val Gly Leu Pro Leu Gly 245 250 255Arg Leu Arg Asp Ile Gln Val Ala Thr Pro Glu Glu Ser Gly Arg Leu 260 265 270Ile Val Asp Ile Leu Lys Asn Arg Gly Pro Asp Ser Ala Val His Ile 275 280 285Ala Ala Phe Asn Ala Gly Ala Ala Leu Tyr Val Ala Glu Gln Ala Ala 290 295 300Asp Leu Lys Glu Gly Val Ala Leu Ala Leu Glu Ala Val Ala Gly Gly305 310 315 320Glu Ala Leu Asn Gln Leu Glu Arg Leu Lys Gln Lys Glu Glu Glu Leu 325 330 335Tyr Ala71759DNABacillus licheniformis DSM 13gene(1)..(759)trpC - indol-3-glycerol phosphate synthase - BLi02401 71atg ctt aat caa att att gat aga aaa cga gaa gac atc cta aag atc 48Met Leu Asn Gln Ile Ile Asp Arg Lys Arg Glu Asp Ile Leu Lys Ile1 5 10 15aag ctg tcg gaa gat tta aaa ctg cca aag cgg tca ttt aaa aaa gcg 96Lys Leu Ser Glu Asp Leu Lys Leu Pro Lys Arg Ser Phe Lys Lys Ala 20 25 30ctc ctt tct ccg aac cgg ttt gtc gct tta att gcg gag gtg aaa aaa 144Leu Leu Ser Pro Asn Arg Phe Val Ala Leu Ile Ala Glu Val Lys Lys 35 40 45gct tct ccg tca aaa ggc gtg att cag gag ggt ttt gag ccc gtc aag 192Ala Ser Pro Ser Lys Gly Val Ile Gln Glu Gly Phe Glu Pro Val Lys 50 55 60atc gca aaa caa tac gaa cag gct aaa gcc gat tgt ttg tct gtt ctg 240Ile Ala Lys Gln Tyr Glu Gln Ala Lys Ala Asp Cys Leu Ser Val Leu65 70 75 80aca gat aca cct ttt ttt caa ggg aag aac agt tat ttg tca gat gtg 288Thr Asp Thr Pro Phe Phe Gln Gly Lys Asn Ser Tyr Leu Ser Asp Val 85 90 95aaa cgc tcg gtt tcc ctg cct gtg ctg cgc aag gat ttt atc att gat 336Lys Arg Ser Val Ser Leu Pro Val Leu Arg Lys Asp Phe Ile Ile Asp 100 105 110tcc att cag gtt gaa gaa gcg gac cgc atc gga gcg gat gcg atc ctg 384Ser Ile Gln Val Glu Glu Ala Asp Arg Ile Gly Ala Asp Ala Ile Leu 115 120 125ctg atc ggt gaa gcg ctt gaa ccg caa aag ctg ttt gaa ctc tat cag 432Leu Ile Gly Glu Ala Leu Glu Pro Gln Lys Leu Phe Glu Leu Tyr Gln 130 135 140caa gcc gtt gaa aaa ggc atg gat gtg ctc gtc gaa gta cat ggc gaa 480Gln Ala Val Glu Lys Gly Met Asp Val Leu Val Glu Val His Gly Glu145 150 155 160gag acg ctt gag ggc att tta aat gtc ttt aca ccc gaa atc atc ggc 528Glu Thr Leu Glu Gly Ile Leu Asn Val Phe Thr Pro Glu Ile Ile Gly 165 170 175gta aac aat cga aat cta aaa aac ttt gaa acg acc gtc gga caa acg 576Val Asn Asn Arg Asn Leu Lys Asn Phe Glu Thr Thr Val Gly Gln Thr 180 185 190gag cga atg gcg aaa ctc gtt ccg ccg ggg acc gtt ctc atc agc gaa 624Glu Arg Met Ala Lys Leu Val Pro Pro Gly Thr Val Leu Ile Ser Glu 195 200 205agc ggc atc gga aaa tcg gaa gac ctg act ttt gta aaa acg tgc gga 672Ser Gly Ile Gly Lys Ser Glu Asp Leu Thr Phe Val Lys Thr Cys Gly 210 215 220gcc caa gcc gtg ctt gtc ggc gaa tcg ctg atg aga gag acg tca cag 720Ala Gln Ala Val Leu Val Gly Glu Ser Leu Met Arg Glu Thr Ser Gln225 230 235 240ctg aag gct gtc tat gcg tta ttc gga gag gac ggc tga 759Leu Lys Ala Val Tyr Ala Leu Phe Gly Glu Asp Gly 245 25072252PRTBacillus licheniformis DSM 13 72Met Leu Asn Gln Ile Ile Asp Arg Lys Arg Glu Asp Ile Leu Lys Ile1 5 10 15Lys Leu Ser Glu Asp Leu Lys Leu Pro Lys Arg Ser Phe Lys Lys Ala 20 25 30Leu Leu Ser Pro Asn Arg Phe Val Ala Leu Ile Ala Glu Val Lys Lys 35 40 45Ala Ser Pro Ser Lys Gly Val Ile Gln Glu Gly Phe Glu Pro Val Lys 50 55 60Ile Ala Lys Gln Tyr Glu Gln Ala Lys Ala Asp Cys Leu Ser Val Leu65 70 75 80Thr Asp Thr Pro Phe Phe Gln Gly Lys Asn Ser Tyr Leu Ser Asp Val 85 90 95Lys Arg Ser Val Ser Leu Pro Val Leu Arg Lys Asp

Phe Ile Ile Asp 100 105 110Ser Ile Gln Val Glu Glu Ala Asp Arg Ile Gly Ala Asp Ala Ile Leu 115 120 125Leu Ile Gly Glu Ala Leu Glu Pro Gln Lys Leu Phe Glu Leu Tyr Gln 130 135 140Gln Ala Val Glu Lys Gly Met Asp Val Leu Val Glu Val His Gly Glu145 150 155 160Glu Thr Leu Glu Gly Ile Leu Asn Val Phe Thr Pro Glu Ile Ile Gly 165 170 175Val Asn Asn Arg Asn Leu Lys Asn Phe Glu Thr Thr Val Gly Gln Thr 180 185 190Glu Arg Met Ala Lys Leu Val Pro Pro Gly Thr Val Leu Ile Ser Glu 195 200 205Ser Gly Ile Gly Lys Ser Glu Asp Leu Thr Phe Val Lys Thr Cys Gly 210 215 220Ala Gln Ala Val Leu Val Gly Glu Ser Leu Met Arg Glu Thr Ser Gln225 230 235 240Leu Lys Ala Val Tyr Ala Leu Phe Gly Glu Asp Gly 245 25073657DNABacillus licheniformis DSM 13gene(1)..(657)trpF - phosphoribosyl anthranilate isomerase - BLi02400 73atg aag aaa ccg agt cta aaa tat tgc ggg att cat tca ttg gaa gat 48Met Lys Lys Pro Ser Leu Lys Tyr Cys Gly Ile His Ser Leu Glu Asp1 5 10 15tta aag gtg aca gcc cgc tcg aaa gcc gat tat ctc gga ttt att ttc 96Leu Lys Val Thr Ala Arg Ser Lys Ala Asp Tyr Leu Gly Phe Ile Phe 20 25 30gcc gaa agc aaa cgg tcg gta gac ccg gcg gat gtg aaa cgg tgg tgc 144Ala Glu Ser Lys Arg Ser Val Asp Pro Ala Asp Val Lys Arg Trp Cys 35 40 45ggc gaa acg gat acg cca ggt aaa aag ctt gtc ggc gtt ttt gtt aat 192Gly Glu Thr Asp Thr Pro Gly Lys Lys Leu Val Gly Val Phe Val Asn 50 55 60gaa aac atg agc cga atg gct gaa atc gta aaa gat gca ggc ttg gac 240Glu Asn Met Ser Arg Met Ala Glu Ile Val Lys Asp Ala Gly Leu Asp65 70 75 80gtg atc cag ctt cac gga gat gaa acg gcg gca gac atc aag cgg ctt 288Val Ile Gln Leu His Gly Asp Glu Thr Ala Ala Asp Ile Lys Arg Leu 85 90 95aaa agc atg aca gac tgc gag ata tgg aag gcg ctc cct cac ggg aat 336Lys Ser Met Thr Asp Cys Glu Ile Trp Lys Ala Leu Pro His Gly Asn 100 105 110gat acc gta cag agc atg gct tct ttc gca cct tac atc gac ggg tat 384Asp Thr Val Gln Ser Met Ala Ser Phe Ala Pro Tyr Ile Asp Gly Tyr 115 120 125gtc atc gac tcg tct gtc aaa ggg atg aga ggg gga acg gga gtt tcg 432Val Ile Asp Ser Ser Val Lys Gly Met Arg Gly Gly Thr Gly Val Ser 130 135 140ttt tcg tgg gac agc gtc ccg ctg tat ata gat gca gcc caa aga gaa 480Phe Ser Trp Asp Ser Val Pro Leu Tyr Ile Asp Ala Ala Gln Arg Glu145 150 155 160ggc aag cgc ctg ttt atc gcc gga ggc gtg aac ccg gac aca atc gca 528Gly Lys Arg Leu Phe Ile Ala Gly Gly Val Asn Pro Asp Thr Ile Ala 165 170 175gat ctt tta aag cgg cgt ccg ccg gga atc gac ttg gca agc gga atc 576Asp Leu Leu Lys Arg Arg Pro Pro Gly Ile Asp Leu Ala Ser Gly Ile 180 185 190gaa gaa aga ggc aga aaa agc gaa aag ctg atc agc ctt tta gaa gaa 624Glu Glu Arg Gly Arg Lys Ser Glu Lys Leu Ile Ser Leu Leu Glu Glu 195 200 205agg atg ttc gaa cat gta ttc ata tcc gaa tga 657Arg Met Phe Glu His Val Phe Ile Ser Glu 210 21574218PRTBacillus licheniformis DSM 13 74Met Lys Lys Pro Ser Leu Lys Tyr Cys Gly Ile His Ser Leu Glu Asp1 5 10 15Leu Lys Val Thr Ala Arg Ser Lys Ala Asp Tyr Leu Gly Phe Ile Phe 20 25 30Ala Glu Ser Lys Arg Ser Val Asp Pro Ala Asp Val Lys Arg Trp Cys 35 40 45Gly Glu Thr Asp Thr Pro Gly Lys Lys Leu Val Gly Val Phe Val Asn 50 55 60Glu Asn Met Ser Arg Met Ala Glu Ile Val Lys Asp Ala Gly Leu Asp65 70 75 80Val Ile Gln Leu His Gly Asp Glu Thr Ala Ala Asp Ile Lys Arg Leu 85 90 95Lys Ser Met Thr Asp Cys Glu Ile Trp Lys Ala Leu Pro His Gly Asn 100 105 110Asp Thr Val Gln Ser Met Ala Ser Phe Ala Pro Tyr Ile Asp Gly Tyr 115 120 125Val Ile Asp Ser Ser Val Lys Gly Met Arg Gly Gly Thr Gly Val Ser 130 135 140Phe Ser Trp Asp Ser Val Pro Leu Tyr Ile Asp Ala Ala Gln Arg Glu145 150 155 160Gly Lys Arg Leu Phe Ile Ala Gly Gly Val Asn Pro Asp Thr Ile Ala 165 170 175Asp Leu Leu Lys Arg Arg Pro Pro Gly Ile Asp Leu Ala Ser Gly Ile 180 185 190Glu Glu Arg Gly Arg Lys Ser Glu Lys Leu Ile Ser Leu Leu Glu Glu 195 200 205Arg Met Phe Glu His Val Phe Ile Ser Glu 210 215751227DNABacillus licheniformis DSM 13gene(1)..(1227)trpB - tryptophan synthase (beta subunit) - BLi02399 75atg tat tca tat ccg aat gaa caa ggc agg ttt ggt gat ttt ggc ggg 48Met Tyr Ser Tyr Pro Asn Glu Gln Gly Arg Phe Gly Asp Phe Gly Gly1 5 10 15aaa ttc gtc ccg gaa aca ctc atg cag cct ttg gaa gaa atc gaa aaa 96Lys Phe Val Pro Glu Thr Leu Met Gln Pro Leu Glu Glu Ile Glu Lys 20 25 30gcg ttt ctt gaa ctg aaa gac gac cct tca ttt caa aag gat tat caa 144Ala Phe Leu Glu Leu Lys Asp Asp Pro Ser Phe Gln Lys Asp Tyr Gln 35 40 45aag ctt tta aat gac tac tca ggc agg ccg acg gct ttg acg ttc gca 192Lys Leu Leu Asn Asp Tyr Ser Gly Arg Pro Thr Ala Leu Thr Phe Ala 50 55 60gac cag ctc act aag cag ctg ggc gga gcg aag att tac tta aaa aga 240Asp Gln Leu Thr Lys Gln Leu Gly Gly Ala Lys Ile Tyr Leu Lys Arg65 70 75 80gaa gat tta aac cat aca ggc gcc cac aaa atc aac aat gcc ctc ggc 288Glu Asp Leu Asn His Thr Gly Ala His Lys Ile Asn Asn Ala Leu Gly 85 90 95cag gcg ctc ctg gcc aag aaa atg gga aaa aca aag ctc att gct gaa 336Gln Ala Leu Leu Ala Lys Lys Met Gly Lys Thr Lys Leu Ile Ala Glu 100 105 110acc ggt gca ggc cag cac ggt gtg gcg gcg gcg act gtc gcc gcg aag 384Thr Gly Ala Gly Gln His Gly Val Ala Ala Ala Thr Val Ala Ala Lys 115 120 125ttc ggc tta tca tgc atc gtc ttc atg ggg gag gaa gat gtt gcc aga 432Phe Gly Leu Ser Cys Ile Val Phe Met Gly Glu Glu Asp Val Ala Arg 130 135 140caa tcg ctg aac gtg ttc cgg atg aag ctc ttg gga gcg gag gtc gtc 480Gln Ser Leu Asn Val Phe Arg Met Lys Leu Leu Gly Ala Glu Val Val145 150 155 160ccg gtg tca agc gga aac ggg aca tta aaa gat gct aca aac gaa gcg 528Pro Val Ser Ser Gly Asn Gly Thr Leu Lys Asp Ala Thr Asn Glu Ala 165 170 175atc cgc tac tgg gtt cag cat tgc cag gac cat ttt tac atg atc ggt 576Ile Arg Tyr Trp Val Gln His Cys Gln Asp His Phe Tyr Met Ile Gly 180 185 190tcc gtc gtc ggg ccg cat ccg tat cct tat atc gtc cgc gaa ttt caa 624Ser Val Val Gly Pro His Pro Tyr Pro Tyr Ile Val Arg Glu Phe Gln 195 200 205cgg atg atc ggc gaa gag gcg aaa gcg cag ttt tat caa ttg gag caa 672Arg Met Ile Gly Glu Glu Ala Lys Ala Gln Phe Tyr Gln Leu Glu Gln 210 215 220agc ctc ccc gat aaa gtt gtc gcc tgt gtc gga ggc gga agc aat gcg 720Ser Leu Pro Asp Lys Val Val Ala Cys Val Gly Gly Gly Ser Asn Ala225 230 235 240atc ggg atg ttc agc gcg ttc att gaa gaa act gtt gaa ctg atc gga 768Ile Gly Met Phe Ser Ala Phe Ile Glu Glu Thr Val Glu Leu Ile Gly 245 250 255gtc gaa gct gcg gga aaa ggt gtt gac aca cct ctt cat gcg gcc acg 816Val Glu Ala Ala Gly Lys Gly Val Asp Thr Pro Leu His Ala Ala Thr 260 265 270atc aca aag ggt aca aaa ggc gtc atc cac ggt tct ttg acc tac ttg 864Ile Thr Lys Gly Thr Lys Gly Val Ile His Gly Ser Leu Thr Tyr Leu 275 280 285att caa gac gag tac gga cag atc atc gag cct tat tcc atc tca gcc 912Ile Gln Asp Glu Tyr Gly Gln Ile Ile Glu Pro Tyr Ser Ile Ser Ala 290 295 300gga ctt gat tat ccg gga gtc ggc ccc gag cac gcc cat ctt cat caa 960Gly Leu Asp Tyr Pro Gly Val Gly Pro Glu His Ala His Leu His Gln305 310 315 320aca ggg cgc gtc acg tat gaa tct gtc aca gat gat gaa gcg gtc tcg 1008Thr Gly Arg Val Thr Tyr Glu Ser Val Thr Asp Asp Glu Ala Val Ser 325 330 335gca tta agg ctg ctg aca gag act gag ggc att ttg ccg gct att gaa 1056Ala Leu Arg Leu Leu Thr Glu Thr Glu Gly Ile Leu Pro Ala Ile Glu 340 345 350tcc gcc cat gca ttg gca aaa gca ttt gaa atc gct aaa gaa ctg ccg 1104Ser Ala His Ala Leu Ala Lys Ala Phe Glu Ile Ala Lys Glu Leu Pro 355 360 365aga gag aaa agc gtt ctc gtc tgc ctt tca ggc agg gga gac aaa gac 1152Arg Glu Lys Ser Val Leu Val Cys Leu Ser Gly Arg Gly Asp Lys Asp 370 375 380gtc cat acc ttg atg aag gtg ttg gaa gaa gag gtg aat caa gat gtt 1200Val His Thr Leu Met Lys Val Leu Glu Glu Glu Val Asn Gln Asp Val385 390 395 400caa tct gaa aaa aca gcc gtc cga taa 1227Gln Ser Glu Lys Thr Ala Val Arg 40576408PRTBacillus licheniformis DSM 13 76Met Tyr Ser Tyr Pro Asn Glu Gln Gly Arg Phe Gly Asp Phe Gly Gly1 5 10 15Lys Phe Val Pro Glu Thr Leu Met Gln Pro Leu Glu Glu Ile Glu Lys 20 25 30Ala Phe Leu Glu Leu Lys Asp Asp Pro Ser Phe Gln Lys Asp Tyr Gln 35 40 45Lys Leu Leu Asn Asp Tyr Ser Gly Arg Pro Thr Ala Leu Thr Phe Ala 50 55 60Asp Gln Leu Thr Lys Gln Leu Gly Gly Ala Lys Ile Tyr Leu Lys Arg65 70 75 80Glu Asp Leu Asn His Thr Gly Ala His Lys Ile Asn Asn Ala Leu Gly 85 90 95Gln Ala Leu Leu Ala Lys Lys Met Gly Lys Thr Lys Leu Ile Ala Glu 100 105 110Thr Gly Ala Gly Gln His Gly Val Ala Ala Ala Thr Val Ala Ala Lys 115 120 125Phe Gly Leu Ser Cys Ile Val Phe Met Gly Glu Glu Asp Val Ala Arg 130 135 140Gln Ser Leu Asn Val Phe Arg Met Lys Leu Leu Gly Ala Glu Val Val145 150 155 160Pro Val Ser Ser Gly Asn Gly Thr Leu Lys Asp Ala Thr Asn Glu Ala 165 170 175Ile Arg Tyr Trp Val Gln His Cys Gln Asp His Phe Tyr Met Ile Gly 180 185 190Ser Val Val Gly Pro His Pro Tyr Pro Tyr Ile Val Arg Glu Phe Gln 195 200 205Arg Met Ile Gly Glu Glu Ala Lys Ala Gln Phe Tyr Gln Leu Glu Gln 210 215 220Ser Leu Pro Asp Lys Val Val Ala Cys Val Gly Gly Gly Ser Asn Ala225 230 235 240Ile Gly Met Phe Ser Ala Phe Ile Glu Glu Thr Val Glu Leu Ile Gly 245 250 255Val Glu Ala Ala Gly Lys Gly Val Asp Thr Pro Leu His Ala Ala Thr 260 265 270Ile Thr Lys Gly Thr Lys Gly Val Ile His Gly Ser Leu Thr Tyr Leu 275 280 285Ile Gln Asp Glu Tyr Gly Gln Ile Ile Glu Pro Tyr Ser Ile Ser Ala 290 295 300Gly Leu Asp Tyr Pro Gly Val Gly Pro Glu His Ala His Leu His Gln305 310 315 320Thr Gly Arg Val Thr Tyr Glu Ser Val Thr Asp Asp Glu Ala Val Ser 325 330 335Ala Leu Arg Leu Leu Thr Glu Thr Glu Gly Ile Leu Pro Ala Ile Glu 340 345 350Ser Ala His Ala Leu Ala Lys Ala Phe Glu Ile Ala Lys Glu Leu Pro 355 360 365Arg Glu Lys Ser Val Leu Val Cys Leu Ser Gly Arg Gly Asp Lys Asp 370 375 380Val His Thr Leu Met Lys Val Leu Glu Glu Glu Val Asn Gln Asp Val385 390 395 400Gln Ser Glu Lys Thr Ala Val Arg 40577210DNABacillus licheniformis DSM 13gene(1)..(210)yppF - unknown function - BLi02362 77atg cac cat atg aac gta tca tac tta aga aat tgt ttt gcc gag atg 48Met His His Met Asn Val Ser Tyr Leu Arg Asn Cys Phe Ala Glu Met1 5 10 15aaa cag tat gag aca gac tgc atg aac aaa ctg atg gat ttc gct aag 96Lys Gln Tyr Glu Thr Asp Cys Met Asn Lys Leu Met Asp Phe Ala Lys 20 25 30ttt ttg tat atc cag gga cat ctg aca tta aac gaa ttt cgc aca agt 144Phe Leu Tyr Ile Gln Gly His Leu Thr Leu Asn Glu Phe Arg Thr Ser 35 40 45atg aaa gtt ctt gaa gcg aat ggc gca cag cac cct gct tat gat atg 192Met Lys Val Leu Glu Ala Asn Gly Ala Gln His Pro Ala Tyr Asp Met 50 55 60aat acg gac gcc agc taa 210Asn Thr Asp Ala Ser657869PRTBacillus licheniformis DSM 13 78Met His His Met Asn Val Ser Tyr Leu Arg Asn Cys Phe Ala Glu Met1 5 10 15Lys Gln Tyr Glu Thr Asp Cys Met Asn Lys Leu Met Asp Phe Ala Lys 20 25 30Phe Leu Tyr Ile Gln Gly His Leu Thr Leu Asn Glu Phe Arg Thr Ser 35 40 45Met Lys Val Leu Glu Ala Asn Gly Ala Gln His Pro Ala Tyr Asp Met 50 55 60Asn Thr Asp Ala Ser6579711DNABacillus licheniformis DSM 13gene(1)..(711)ydfS - unknown function - similar to unknown function proteins - BLi01489 79atg agt gaa cat att gaa gta atc ctt cga tcg att ctt gcc ttt ggt 48Met Ser Glu His Ile Glu Val Ile Leu Arg Ser Ile Leu Ala Phe Gly1 5 10 15att tta ttg gta gga tca aga ttg ttg gga aaa caa aca att tca caa 96Ile Leu Leu Val Gly Ser Arg Leu Leu Gly Lys Gln Thr Ile Ser Gln 20 25 30atg aat atc ttt gat ttt att gca tca att aca tta ggc gct att act 144Met Asn Ile Phe Asp Phe Ile Ala Ser Ile Thr Leu Gly Ala Ile Thr 35 40 45gca aat ctg gct ttt aat acc tct ctt aaa att cat cac atg gta ctg 192Ala Asn Leu Ala Phe Asn Thr Ser Leu Lys Ile His His Met Val Leu 50 55 60gct ttt ttt ata ttt gtt ttt gtg atc tta gcc act gcg tat ata tcc 240Ala Phe Phe Ile Phe Val Phe Val Ile Leu Ala Thr Ala Tyr Ile Ser65 70 75 80ctt tgg agt caa aaa ggc cgc aaa ttt ttt gcc ggt gat cca act gtt 288Leu Trp Ser Gln Lys Gly Arg Lys Phe Phe Ala Gly Asp Pro Thr Val 85 90 95gtg att caa aat gga aaa gtt ctt gaa cac aac atg cga aaa atg cgt 336Val Ile Gln Asn Gly Lys Val Leu Glu His Asn Met Arg Lys Met Arg 100 105 110tat aca att gat tat ttg aat caa caa ctt aga gaa aag aat gta ttc 384Tyr Thr Ile Asp Tyr Leu Asn Gln Gln Leu Arg Glu Lys Asn Val Phe 115 120 125aaa atc gaa gaa gta ctc tat gcc att gtt gaa aca aat ggg acc tta 432Lys Ile Glu Glu Val Leu Tyr Ala Ile Val Glu Thr Asn Gly Thr Leu 130 135 140aca gta tta aaa aaa cca caa ttc cga aat gtg aca aaa aag gat ttg 480Thr Val Leu Lys Lys Pro Gln Phe Arg Asn Val Thr Lys Lys Asp Leu145 150 155 160tgg gtt gcc act acg cca gaa gga aaa ctc ccg att gaa tta atc atg 528Trp Val Ala Thr Thr Pro Glu Gly Lys Leu Pro Ile Glu Leu Ile Met 165 170 175gat ggg aaa atg aag aat gaa aac ttg aaa gat aat caa ctc act cag 576Asp Gly Lys Met Lys Asn Glu Asn Leu Lys Asp Asn Gln Leu Thr Gln 180 185 190gca tgg tta caa tct gag atg aaa aag cgt ggt cta tcc aca gat gaa 624Ala Trp Leu Gln Ser Glu Met Lys Lys Arg Gly Leu Ser Thr Asp Glu 195 200 205gtt ttt tat gca gta tta tca cct gat gga aat atc tat ata gat aca 672Val Phe Tyr Ala Val Leu Ser Pro Asp Gly Asn Ile Tyr Ile Asp Thr 210 215 220tat gaa gat cat ata cat tct tct ttt gat aaa gaa taa 711Tyr Glu Asp His Ile His Ser Ser Phe Asp Lys Glu225 230 23580236PRTBacillus licheniformis DSM 13 80Met Ser Glu His Ile Glu Val Ile Leu Arg Ser Ile Leu Ala Phe Gly1 5 10 15Ile Leu Leu Val Gly Ser Arg Leu Leu Gly Lys Gln Thr Ile Ser Gln 20 25 30Met Asn Ile Phe Asp Phe Ile Ala Ser Ile Thr Leu Gly Ala Ile Thr 35 40

45Ala Asn Leu Ala Phe Asn Thr Ser Leu Lys Ile His His Met Val Leu 50 55 60Ala Phe Phe Ile Phe Val Phe Val Ile Leu Ala Thr Ala Tyr Ile Ser65 70 75 80Leu Trp Ser Gln Lys Gly Arg Lys Phe Phe Ala Gly Asp Pro Thr Val 85 90 95Val Ile Gln Asn Gly Lys Val Leu Glu His Asn Met Arg Lys Met Arg 100 105 110Tyr Thr Ile Asp Tyr Leu Asn Gln Gln Leu Arg Glu Lys Asn Val Phe 115 120 125Lys Ile Glu Glu Val Leu Tyr Ala Ile Val Glu Thr Asn Gly Thr Leu 130 135 140Thr Val Leu Lys Lys Pro Gln Phe Arg Asn Val Thr Lys Lys Asp Leu145 150 155 160Trp Val Ala Thr Thr Pro Glu Gly Lys Leu Pro Ile Glu Leu Ile Met 165 170 175Asp Gly Lys Met Lys Asn Glu Asn Leu Lys Asp Asn Gln Leu Thr Gln 180 185 190Ala Trp Leu Gln Ser Glu Met Lys Lys Arg Gly Leu Ser Thr Asp Glu 195 200 205Val Phe Tyr Ala Val Leu Ser Pro Asp Gly Asn Ile Tyr Ile Asp Thr 210 215 220Tyr Glu Asp His Ile His Ser Ser Phe Asp Lys Glu225 230 23581852DNABacillus licheniformis DSM 13gene(1)..(852)hypothetical protein - unknown function - BLi01488 81atg cct ccc gac aga aaa cta att aat gac att cac aag gca att aac 48Met Pro Pro Asp Arg Lys Leu Ile Asn Asp Ile His Lys Ala Ile Asn1 5 10 15gga gag cat agc gcc att ctc tgc tat aag caa ata gcc gaa aat gcc 96Gly Glu His Ser Ala Ile Leu Cys Tyr Lys Gln Ile Ala Glu Asn Ala 20 25 30ccc aca gaa gat ata aga aaa caa ata ctt gaa att aga caa gat gag 144Pro Thr Glu Asp Ile Arg Lys Gln Ile Leu Glu Ile Arg Gln Asp Glu 35 40 45caa cga cat ttt gaa gag ttt agc aga att ttc act aac ctg aca ggc 192Gln Arg His Phe Glu Glu Phe Ser Arg Ile Phe Thr Asn Leu Thr Gly 50 55 60aaa aag cca atc ccc ata atc ata gaa gaa tgc cct aaa aag tat aga 240Lys Lys Pro Ile Pro Ile Ile Ile Glu Glu Cys Pro Lys Lys Tyr Arg65 70 75 80aca agt cta gag tct gct ttt aaa gat gag cag gag aca gtc gat ttt 288Thr Ser Leu Glu Ser Ala Phe Lys Asp Glu Gln Glu Thr Val Asp Phe 85 90 95tat tta gat tta gca gac cga ata caa aat cag tca att aaa aag gtt 336Tyr Leu Asp Leu Ala Asp Arg Ile Gln Asn Gln Ser Ile Lys Lys Val 100 105 110ttt cag cgt gca gca gct gat gaa caa aat cat gct gta tgg ttc tta 384Phe Gln Arg Ala Ala Ala Asp Glu Gln Asn His Ala Val Trp Phe Leu 115 120 125ttt ttt tta aat aca tcg tct aca caa caa aca cgt caa gtt aat tat 432Phe Phe Leu Asn Thr Ser Ser Thr Gln Gln Thr Arg Gln Val Asn Tyr 130 135 140ggg gca aaa gga gcg tta aat tcc ggt agg ctc acc tta cct caa atg 480Gly Ala Lys Gly Ala Leu Asn Ser Gly Arg Leu Thr Leu Pro Gln Met145 150 155 160ctt acc ttt gcc att cag gat gag tac tta gct caa gca aga tac aat 528Leu Thr Phe Ala Ile Gln Asp Glu Tyr Leu Ala Gln Ala Arg Tyr Asn 165 170 175gaa att cta aga aac ttc ggg gat att cga acc ttt gtc caa att aaa 576Glu Ile Leu Arg Asn Phe Gly Asp Ile Arg Thr Phe Val Gln Ile Lys 180 185 190gaa gcc gaa tta cgg cat atc aat gca tta ctt tct ctt ttt gaa cga 624Glu Ala Glu Leu Arg His Ile Asn Ala Leu Leu Ser Leu Phe Glu Arg 195 200 205tac caa gta cag ata cct aaa gac acc tct caa tta tat gtc act aca 672Tyr Gln Val Gln Ile Pro Lys Asp Thr Ser Gln Leu Tyr Val Thr Thr 210 215 220cca gta acc ctt aag gat gca tat gct tct ggg gta caa gga gaa att 720Pro Val Thr Leu Lys Asp Ala Tyr Ala Ser Gly Val Gln Gly Glu Ile225 230 235 240gat aat atc gcc atg tat aac aag ttt cta act ttt gat ctt cca gca 768Asp Asn Ile Ala Met Tyr Asn Lys Phe Leu Thr Phe Asp Leu Pro Ala 245 250 255gat gca aga aca gtt ttt act caa tta cgc aac gca tca cta aat cat 816Asp Ala Arg Thr Val Phe Thr Gln Leu Arg Asn Ala Ser Leu Asn His 260 265 270tta gag gct ttc cga aga ggt ctt gag aga aat taa 852Leu Glu Ala Phe Arg Arg Gly Leu Glu Arg Asn 275 28082283PRTBacillus licheniformis DSM 13 82Met Pro Pro Asp Arg Lys Leu Ile Asn Asp Ile His Lys Ala Ile Asn1 5 10 15Gly Glu His Ser Ala Ile Leu Cys Tyr Lys Gln Ile Ala Glu Asn Ala 20 25 30Pro Thr Glu Asp Ile Arg Lys Gln Ile Leu Glu Ile Arg Gln Asp Glu 35 40 45Gln Arg His Phe Glu Glu Phe Ser Arg Ile Phe Thr Asn Leu Thr Gly 50 55 60Lys Lys Pro Ile Pro Ile Ile Ile Glu Glu Cys Pro Lys Lys Tyr Arg65 70 75 80Thr Ser Leu Glu Ser Ala Phe Lys Asp Glu Gln Glu Thr Val Asp Phe 85 90 95Tyr Leu Asp Leu Ala Asp Arg Ile Gln Asn Gln Ser Ile Lys Lys Val 100 105 110Phe Gln Arg Ala Ala Ala Asp Glu Gln Asn His Ala Val Trp Phe Leu 115 120 125Phe Phe Leu Asn Thr Ser Ser Thr Gln Gln Thr Arg Gln Val Asn Tyr 130 135 140Gly Ala Lys Gly Ala Leu Asn Ser Gly Arg Leu Thr Leu Pro Gln Met145 150 155 160Leu Thr Phe Ala Ile Gln Asp Glu Tyr Leu Ala Gln Ala Arg Tyr Asn 165 170 175Glu Ile Leu Arg Asn Phe Gly Asp Ile Arg Thr Phe Val Gln Ile Lys 180 185 190Glu Ala Glu Leu Arg His Ile Asn Ala Leu Leu Ser Leu Phe Glu Arg 195 200 205Tyr Gln Val Gln Ile Pro Lys Asp Thr Ser Gln Leu Tyr Val Thr Thr 210 215 220Pro Val Thr Leu Lys Asp Ala Tyr Ala Ser Gly Val Gln Gly Glu Ile225 230 235 240Asp Asn Ile Ala Met Tyr Asn Lys Phe Leu Thr Phe Asp Leu Pro Ala 245 250 255Asp Ala Arg Thr Val Phe Thr Gln Leu Arg Asn Ala Ser Leu Asn His 260 265 270Leu Glu Ala Phe Arg Arg Gly Leu Glu Arg Asn 275 28083594DNABacillus licheniformis DSM 13gene(1)..(594)yrkC - unknown function - similar to unknown function proteins - BLi01487 83atg tat tat gtt cct cat atg tat cca tat caa tat tct tat cct tat 48Met Tyr Tyr Val Pro His Met Tyr Pro Tyr Gln Tyr Ser Tyr Pro Tyr1 5 10 15tat gtt aat gtg cca atg tac aac tgt aga aga cag tca gtt cac cgg 96Tyr Val Asn Val Pro Met Tyr Asn Cys Arg Arg Gln Ser Val His Arg 20 25 30act gtt cct agt gaa gta gag cat gca aac aaa tta agt tct gtt cgc 144Thr Val Pro Ser Glu Val Glu His Ala Asn Lys Leu Ser Ser Val Arg 35 40 45tca tcc aaa ggt agt aac agc att ttg tta aag gat tat gga cca gaa 192Ser Ser Lys Gly Ser Asn Ser Ile Leu Leu Lys Asp Tyr Gly Pro Glu 50 55 60gcg ttt gtg gtt aat atc aat gaa gct acg aag caa aat aat acg tac 240Ala Phe Val Val Asn Ile Asn Glu Ala Thr Lys Gln Asn Asn Thr Tyr65 70 75 80cgt acc gct tta tgg aca gga act cat ttg caa gtc acc tta atg agt 288Arg Thr Ala Leu Trp Thr Gly Thr His Leu Gln Val Thr Leu Met Ser 85 90 95ctc aat gtt gga gaa gat att ggt ttg gaa atg cac cct aac gtt gat 336Leu Asn Val Gly Glu Asp Ile Gly Leu Glu Met His Pro Asn Val Asp 100 105 110caa ttc tta cgt gtt gaa caa ggt cag gga atc gtt caa atg ggc aag 384Gln Phe Leu Arg Val Glu Gln Gly Gln Gly Ile Val Gln Met Gly Lys 115 120 125aat aaa gac cat tta aat ttc aaa aga aat gtc tat gat gat tct gca 432Asn Lys Asp His Leu Asn Phe Lys Arg Asn Val Tyr Asp Asp Ser Ala 130 135 140atc ata ata ccc gct gga aca tgg cac aat cta acc aat aca ggt aga 480Ile Ile Ile Pro Ala Gly Thr Trp His Asn Leu Thr Asn Thr Gly Arg145 150 155 160atc ccg cta aaa cta tac tcc ata tat gct cct cct aac cat cca ttt 528Ile Pro Leu Lys Leu Tyr Ser Ile Tyr Ala Pro Pro Asn His Pro Phe 165 170 175ggt act gtt cat gtt act aaa gga gat gca tta gct tca gaa gaa gaa 576Gly Thr Val His Val Thr Lys Gly Asp Ala Leu Ala Ser Glu Glu Glu 180 185 190tac ggt cat ggc aat tga 594Tyr Gly His Gly Asn 19584197PRTBacillus licheniformis DSM 13 84Met Tyr Tyr Val Pro His Met Tyr Pro Tyr Gln Tyr Ser Tyr Pro Tyr1 5 10 15Tyr Val Asn Val Pro Met Tyr Asn Cys Arg Arg Gln Ser Val His Arg 20 25 30Thr Val Pro Ser Glu Val Glu His Ala Asn Lys Leu Ser Ser Val Arg 35 40 45Ser Ser Lys Gly Ser Asn Ser Ile Leu Leu Lys Asp Tyr Gly Pro Glu 50 55 60Ala Phe Val Val Asn Ile Asn Glu Ala Thr Lys Gln Asn Asn Thr Tyr65 70 75 80Arg Thr Ala Leu Trp Thr Gly Thr His Leu Gln Val Thr Leu Met Ser 85 90 95Leu Asn Val Gly Glu Asp Ile Gly Leu Glu Met His Pro Asn Val Asp 100 105 110Gln Phe Leu Arg Val Glu Gln Gly Gln Gly Ile Val Gln Met Gly Lys 115 120 125Asn Lys Asp His Leu Asn Phe Lys Arg Asn Val Tyr Asp Asp Ser Ala 130 135 140Ile Ile Ile Pro Ala Gly Thr Trp His Asn Leu Thr Asn Thr Gly Arg145 150 155 160Ile Pro Leu Lys Leu Tyr Ser Ile Tyr Ala Pro Pro Asn His Pro Phe 165 170 175Gly Thr Val His Val Thr Lys Gly Asp Ala Leu Ala Ser Glu Glu Glu 180 185 190Tyr Gly His Gly Asn 19585954DNABacillus licheniformis DSM 13gene(1)..(954)putative protein - putative glycosyl hydrolase/lysozyme - BLi01486 85atg gga atc aaa gga atc gac gta tca cac tgg caa ggt aat atc aat 48Met Gly Ile Lys Gly Ile Asp Val Ser His Trp Gln Gly Asn Ile Asn1 5 10 15tgg aag aaa gtt gcg ggg gac ggt att aaa ttc gct ttt atc aaa gca 96Trp Lys Lys Val Ala Gly Asp Gly Ile Lys Phe Ala Phe Ile Lys Ala 20 25 30aca gaa ggg aca aca tta cag gac aat aaa ttt gta acg aat att tca 144Thr Glu Gly Thr Thr Leu Gln Asp Asn Lys Phe Val Thr Asn Ile Ser 35 40 45ggt gct aac gct gtg ggg att aaa acg gga gcc tac cac ttt gca aga 192Gly Ala Asn Ala Val Gly Ile Lys Thr Gly Ala Tyr His Phe Ala Arg 50 55 60ttc ggt tcc aag tca gaa gca ttg gca gag gcc agg ttc ttt ttg tca 240Phe Gly Ser Lys Ser Glu Ala Leu Ala Glu Ala Arg Phe Phe Leu Ser65 70 75 80gtt gca aat aag gtc cat ctc acg tat ccg ctt gtg ctt gat ctt gaa 288Val Ala Asn Lys Val His Leu Thr Tyr Pro Leu Val Leu Asp Leu Glu 85 90 95gtt aat cag cgg aat gtc agt aaa tca gtt ttg aca gat gca gca gtg 336Val Asn Gln Arg Asn Val Ser Lys Ser Val Leu Thr Asp Ala Ala Val 100 105 110gcc ttt tta cgg gaa gtt gaa aaa gct ggt tac ttc gcc atg ata tac 384Ala Phe Leu Arg Glu Val Glu Lys Ala Gly Tyr Phe Ala Met Ile Tyr 115 120 125agc ggt aag tct ttc ctt gag aat tgc ctt gac gaa tcc aag ctg aag 432Ser Gly Lys Ser Phe Leu Glu Asn Cys Leu Asp Glu Ser Lys Leu Lys 130 135 140cca ttt gca tta tgg gtt gct cgt tat aac aac aca ctt ggc cgt cat 480Pro Phe Ala Leu Trp Val Ala Arg Tyr Asn Asn Thr Leu Gly Arg His145 150 155 160gca gat atc tgg cag tat tct gat tgt gga agg gtc gcc ggt att tct 528Ala Asp Ile Trp Gln Tyr Ser Asp Cys Gly Arg Val Ala Gly Ile Ser 165 170 175ggg aat gtt gat atg aat att tgt tat cgt gac ggg tta cgg gct cag 576Gly Asn Val Asp Met Asn Ile Cys Tyr Arg Asp Gly Leu Arg Ala Gln 180 185 190gta gcc gtg aca act gaa aaa gct gct act gta aaa cct gtt tca aac 624Val Ala Val Thr Thr Glu Lys Ala Ala Thr Val Lys Pro Val Ser Asn 195 200 205aaa aag cca gtt aaa acg gag aca gtt tac aca gtc aaa aaa ggg gac 672Lys Lys Pro Val Lys Thr Glu Thr Val Tyr Thr Val Lys Lys Gly Asp 210 215 220acg ctt tcg gaa atc gca cag aaa aac aac acg acc gtt aaa gct ctc 720Thr Leu Ser Glu Ile Ala Gln Lys Asn Asn Thr Thr Val Lys Ala Leu225 230 235 240caa aat ttg aat aac att aaa gat gcg aat aag att tac gtt ggc cag 768Gln Asn Leu Asn Asn Ile Lys Asp Ala Asn Lys Ile Tyr Val Gly Gln 245 250 255aaa tta aag att agc ggc agt cct tca aca gca tcg aat aaa aag caa 816Lys Leu Lys Ile Ser Gly Ser Pro Ser Thr Ala Ser Asn Lys Lys Gln 260 265 270tat tac acc att aaa tcc ggt gac act ttg tct ggg atc tcg aaa agg 864Tyr Tyr Thr Ile Lys Ser Gly Asp Thr Leu Ser Gly Ile Ser Lys Arg 275 280 285ttt aat aca tcc att aag acg ctg caa gct tgg aat ggt atc aag aat 912Phe Asn Thr Ser Ile Lys Thr Leu Gln Ala Trp Asn Gly Ile Lys Asn 290 295 300gcc aac aag att tac gct ggt cag aaa att cgt gtt aga taa 954Ala Asn Lys Ile Tyr Ala Gly Gln Lys Ile Arg Val Arg305 310 31586317PRTBacillus licheniformis DSM 13 86Met Gly Ile Lys Gly Ile Asp Val Ser His Trp Gln Gly Asn Ile Asn1 5 10 15Trp Lys Lys Val Ala Gly Asp Gly Ile Lys Phe Ala Phe Ile Lys Ala 20 25 30Thr Glu Gly Thr Thr Leu Gln Asp Asn Lys Phe Val Thr Asn Ile Ser 35 40 45Gly Ala Asn Ala Val Gly Ile Lys Thr Gly Ala Tyr His Phe Ala Arg 50 55 60Phe Gly Ser Lys Ser Glu Ala Leu Ala Glu Ala Arg Phe Phe Leu Ser65 70 75 80Val Ala Asn Lys Val His Leu Thr Tyr Pro Leu Val Leu Asp Leu Glu 85 90 95Val Asn Gln Arg Asn Val Ser Lys Ser Val Leu Thr Asp Ala Ala Val 100 105 110Ala Phe Leu Arg Glu Val Glu Lys Ala Gly Tyr Phe Ala Met Ile Tyr 115 120 125Ser Gly Lys Ser Phe Leu Glu Asn Cys Leu Asp Glu Ser Lys Leu Lys 130 135 140Pro Phe Ala Leu Trp Val Ala Arg Tyr Asn Asn Thr Leu Gly Arg His145 150 155 160Ala Asp Ile Trp Gln Tyr Ser Asp Cys Gly Arg Val Ala Gly Ile Ser 165 170 175Gly Asn Val Asp Met Asn Ile Cys Tyr Arg Asp Gly Leu Arg Ala Gln 180 185 190Val Ala Val Thr Thr Glu Lys Ala Ala Thr Val Lys Pro Val Ser Asn 195 200 205Lys Lys Pro Val Lys Thr Glu Thr Val Tyr Thr Val Lys Lys Gly Asp 210 215 220Thr Leu Ser Glu Ile Ala Gln Lys Asn Asn Thr Thr Val Lys Ala Leu225 230 235 240Gln Asn Leu Asn Asn Ile Lys Asp Ala Asn Lys Ile Tyr Val Gly Gln 245 250 255Lys Leu Lys Ile Ser Gly Ser Pro Ser Thr Ala Ser Asn Lys Lys Gln 260 265 270Tyr Tyr Thr Ile Lys Ser Gly Asp Thr Leu Ser Gly Ile Ser Lys Arg 275 280 285Phe Asn Thr Ser Ile Lys Thr Leu Gln Ala Trp Asn Gly Ile Lys Asn 290 295 300Ala Asn Lys Ile Tyr Ala Gly Gln Lys Ile Arg Val Arg305 310 31587474DNABacillus licheniformis DSM 13gene(1)..(474)yncE - unknown function - BLi03150 87atg agt cat aag cga ggc gtt tta ttt gaa atc ccg aac ggg tac ggg 48Met Ser His Lys Arg Gly Val Leu Phe Glu Ile Pro Asn Gly Tyr Gly1 5 10 15cgc ttt ctt ctc gat ata ctg aag ccg gtt gaa ccg gcc gat ttc aat 96Arg Phe Leu Leu Asp Ile Leu Lys Pro Val Glu Pro Ala Asp Phe Asn 20 25 30tgg ctg ctt gcc gat ggc gaa tgc tat ctc gtg gaa aac ggt gaa ttg 144Trp Leu Leu Ala Asp Gly Glu Cys Tyr Leu Val Glu Asn Gly Glu Leu 35 40 45tca gat gaa aat ctt ttt cct caa gat ccg cag gtg atg aca gga gaa 192Ser Asp Glu Asn Leu Phe Pro Gln Asp Pro Gln Val Met Thr Gly Glu 50 55 60gca ttt aaa aaa agg ctg gaa gac gca agg tac tat ctc atc ttc gcc 240Ala Phe Lys Lys Arg Leu Glu Asp Ala Arg Tyr Tyr Leu Ile Phe Ala65

70 75 80gat ctg aag gct ttt ccg aag gat ggc cat ccc gaa gat gtc agc aca 288Asp Leu Lys Ala Phe Pro Lys Asp Gly His Pro Glu Asp Val Ser Thr 85 90 95tat gaa tca tat atg aaa agc agc tgc gag ctg att gtg ctt aca gcc 336Tyr Glu Ser Tyr Met Lys Ser Ser Cys Glu Leu Ile Val Leu Thr Ala 100 105 110gat tcg agt ttt gtc acg ata tac tgc aaa gat caa aaa aac ctg gag 384Asp Ser Ser Phe Val Thr Ile Tyr Cys Lys Asp Gln Lys Asn Leu Glu 115 120 125ctt cta tat caa aat gca aag gac tgc ggt ttc aag cgg att gaa tat 432Leu Leu Tyr Gln Asn Ala Lys Asp Cys Gly Phe Lys Arg Ile Glu Tyr 130 135 140ata acg gat cac aac gat gcc aga aca gga atg cat gtg tag 474Ile Thr Asp His Asn Asp Ala Arg Thr Gly Met His Val145 150 15588157PRTBacillus licheniformis DSM 13 88Met Ser His Lys Arg Gly Val Leu Phe Glu Ile Pro Asn Gly Tyr Gly1 5 10 15Arg Phe Leu Leu Asp Ile Leu Lys Pro Val Glu Pro Ala Asp Phe Asn 20 25 30Trp Leu Leu Ala Asp Gly Glu Cys Tyr Leu Val Glu Asn Gly Glu Leu 35 40 45Ser Asp Glu Asn Leu Phe Pro Gln Asp Pro Gln Val Met Thr Gly Glu 50 55 60Ala Phe Lys Lys Arg Leu Glu Asp Ala Arg Tyr Tyr Leu Ile Phe Ala65 70 75 80Asp Leu Lys Ala Phe Pro Lys Asp Gly His Pro Glu Asp Val Ser Thr 85 90 95Tyr Glu Ser Tyr Met Lys Ser Ser Cys Glu Leu Ile Val Leu Thr Ala 100 105 110Asp Ser Ser Phe Val Thr Ile Tyr Cys Lys Asp Gln Lys Asn Leu Glu 115 120 125Leu Leu Tyr Gln Asn Ala Lys Asp Cys Gly Phe Lys Arg Ile Glu Tyr 130 135 140Ile Thr Asp His Asn Asp Ala Arg Thr Gly Met His Val145 150 155891095DNABacillus licheniformis DSM 13gene(1)..(1095)yvlB - unknown function - similar to unknown function proteins - BLi03752 89atg aag cat gag aaa gaa cga att tta aaa ttg gtc gag gaa gga aag 48Met Lys His Glu Lys Glu Arg Ile Leu Lys Leu Val Glu Glu Gly Lys1 5 10 15ctg tct gca aaa gag gcg ctg atc ctg atc gaa aaa ctt gaa gaa gac 96Leu Ser Ala Lys Glu Ala Leu Ile Leu Ile Glu Lys Leu Glu Glu Asp 20 25 30tac aag gaa acc gag agc aag gtg aca gcc ctg tct gag gat gtc att 144Tyr Lys Glu Thr Glu Ser Lys Val Thr Ala Leu Ser Glu Asp Val Ile 35 40 45gat tcg gag gac ttc tac agc gaa aag aaa aaa gaa ccg aag cct tcg 192Asp Ser Glu Asp Phe Tyr Ser Glu Lys Lys Lys Glu Pro Lys Pro Ser 50 55 60atc ggt tca aaa ctg ttc gaa tgg att gat acc gct gta aaa aaa gta 240Ile Gly Ser Lys Leu Phe Glu Trp Ile Asp Thr Ala Val Lys Lys Val65 70 75 80aaa gaa gca gac ttg gat ttg aat ttc ggg cac tcg tac gat gtc cag 288Lys Glu Ala Asp Leu Asp Leu Asn Phe Gly His Ser Tyr Asp Val Gln 85 90 95cat ata ttc cag ttt aaa gac act gat ttt tcg act ctg gat att cag 336His Ile Phe Gln Phe Lys Asp Thr Asp Phe Ser Thr Leu Asp Ile Gln 100 105 110ctt gcc aac ggc agc gtc aac ctt gta cca tgg agc gat tcc gac atc 384Leu Ala Asn Gly Ser Val Asn Leu Val Pro Trp Ser Asp Ser Asp Ile 115 120 125aga gtc gaa tgc cag gca aaa gtg tac cgg gct gaa aat cag gat gaa 432Arg Val Glu Cys Gln Ala Lys Val Tyr Arg Ala Glu Asn Gln Asp Glu 130 135 140gca agg cag gct ttt ctc cag cat atc gac tgc ggt gtg gaa ggc gat 480Ala Arg Gln Ala Phe Leu Gln His Ile Asp Cys Gly Val Glu Gly Asp145 150 155 160aag ttc atg atc cgt acc gag aaa aaa acg atg aag acc aac ctt acc 528Lys Phe Met Ile Arg Thr Glu Lys Lys Thr Met Lys Thr Asn Leu Thr 165 170 175gtc tat ata ccg gac aaa gag tac ggc aaa atc cgc ttt aaa ctg ttt 576Val Tyr Ile Pro Asp Lys Glu Tyr Gly Lys Ile Arg Phe Lys Leu Phe 180 185 190aac ggc ccg gtc aga ggg gaa cat tta aac gta aaa gag ctg tcc gcc 624Asn Gly Pro Val Arg Gly Glu His Leu Asn Val Lys Glu Leu Ser Ala 195 200 205aaa acg acg aac ggc gtg ctg tcg ttc tct gct ctc acg gca gga aaa 672Lys Thr Thr Asn Gly Val Leu Ser Phe Ser Ala Leu Thr Ala Gly Lys 210 215 220gct tca ttg gaa act gca aat gga caa atc aaa cta gcc gac cat act 720Ala Ser Leu Glu Thr Ala Asn Gly Gln Ile Lys Leu Ala Asp His Thr225 230 235 240tgc gga gag att gaa gcg gaa acc atc aat gga ttg atc gac ctt aaa 768Cys Gly Glu Ile Glu Ala Glu Thr Ile Asn Gly Leu Ile Asp Leu Lys 245 250 255ggt tca tgc gag gcg ctt gac ctg cag agc ttt aac ggc aat atc gcg 816Gly Ser Cys Glu Ala Leu Asp Leu Gln Ser Phe Asn Gly Asn Ile Ala 260 265 270gca aca gtc aaa aat ccg cac tgc cgg acg gca tat gtg aaa aca acg 864Ala Thr Val Lys Asn Pro His Cys Arg Thr Ala Tyr Val Lys Thr Thr 275 280 285act gga agc gta gag ttg aat gtt ccc cgg gac tgc gct gtt aca gcg 912Thr Gly Ser Val Glu Leu Asn Val Pro Arg Asp Cys Ala Val Thr Ala 290 295 300gag ctg aaa agc aat ctc ggt tct ctg tct cat gat ctt tca gat gca 960Glu Leu Lys Ser Asn Leu Gly Ser Leu Ser His Asp Leu Ser Asp Ala305 310 315 320gaa att tta aag gaa aaa aac gag aca atc cag aag gaa atg tac cta 1008Glu Ile Leu Lys Glu Lys Asn Glu Thr Ile Gln Lys Glu Met Tyr Leu 325 330 335aaa gca aat caa aat cat gat cat agc atg acc gtg ttt ttg gaa tcg 1056Lys Ala Asn Gln Asn His Asp His Ser Met Thr Val Phe Leu Glu Ser 340 345 350aag aca gga tcg atc cat tta aac cat aca cag gag tga 1095Lys Thr Gly Ser Ile His Leu Asn His Thr Gln Glu 355 36090364PRTBacillus licheniformis DSM 13 90Met Lys His Glu Lys Glu Arg Ile Leu Lys Leu Val Glu Glu Gly Lys1 5 10 15Leu Ser Ala Lys Glu Ala Leu Ile Leu Ile Glu Lys Leu Glu Glu Asp 20 25 30Tyr Lys Glu Thr Glu Ser Lys Val Thr Ala Leu Ser Glu Asp Val Ile 35 40 45Asp Ser Glu Asp Phe Tyr Ser Glu Lys Lys Lys Glu Pro Lys Pro Ser 50 55 60Ile Gly Ser Lys Leu Phe Glu Trp Ile Asp Thr Ala Val Lys Lys Val65 70 75 80Lys Glu Ala Asp Leu Asp Leu Asn Phe Gly His Ser Tyr Asp Val Gln 85 90 95His Ile Phe Gln Phe Lys Asp Thr Asp Phe Ser Thr Leu Asp Ile Gln 100 105 110Leu Ala Asn Gly Ser Val Asn Leu Val Pro Trp Ser Asp Ser Asp Ile 115 120 125Arg Val Glu Cys Gln Ala Lys Val Tyr Arg Ala Glu Asn Gln Asp Glu 130 135 140Ala Arg Gln Ala Phe Leu Gln His Ile Asp Cys Gly Val Glu Gly Asp145 150 155 160Lys Phe Met Ile Arg Thr Glu Lys Lys Thr Met Lys Thr Asn Leu Thr 165 170 175Val Tyr Ile Pro Asp Lys Glu Tyr Gly Lys Ile Arg Phe Lys Leu Phe 180 185 190Asn Gly Pro Val Arg Gly Glu His Leu Asn Val Lys Glu Leu Ser Ala 195 200 205Lys Thr Thr Asn Gly Val Leu Ser Phe Ser Ala Leu Thr Ala Gly Lys 210 215 220Ala Ser Leu Glu Thr Ala Asn Gly Gln Ile Lys Leu Ala Asp His Thr225 230 235 240Cys Gly Glu Ile Glu Ala Glu Thr Ile Asn Gly Leu Ile Asp Leu Lys 245 250 255Gly Ser Cys Glu Ala Leu Asp Leu Gln Ser Phe Asn Gly Asn Ile Ala 260 265 270Ala Thr Val Lys Asn Pro His Cys Arg Thr Ala Tyr Val Lys Thr Thr 275 280 285Thr Gly Ser Val Glu Leu Asn Val Pro Arg Asp Cys Ala Val Thr Ala 290 295 300Glu Leu Lys Ser Asn Leu Gly Ser Leu Ser His Asp Leu Ser Asp Ala305 310 315 320Glu Ile Leu Lys Glu Lys Asn Glu Thr Ile Gln Lys Glu Met Tyr Leu 325 330 335Lys Ala Asn Gln Asn His Asp His Ser Met Thr Val Phe Leu Glu Ser 340 345 350Lys Thr Gly Ser Ile His Leu Asn His Thr Gln Glu 355 360911182DNABacillus licheniformis DSM 13gene(1)..(1182)putative protein - putative ABC transporter (amino acid permease) - BLi03212 91atg aag gaa aac ctg att ccg gtc act gtt tta agc ggt tat cta ggg 48Met Lys Glu Asn Leu Ile Pro Val Thr Val Leu Ser Gly Tyr Leu Gly1 5 10 15tcc ggg aaa acg acg ctg ctg cag cat gtc ctg cat gaa gcg cac ggc 96Ser Gly Lys Thr Thr Leu Leu Gln His Val Leu His Glu Ala His Gly 20 25 30ttg aaa atc gcc gtg atc gtc aat gat atg agc gag att aat atc gac 144Leu Lys Ile Ala Val Ile Val Asn Asp Met Ser Glu Ile Asn Ile Asp 35 40 45ggc tcc ctc gtg aaa aaa ggt gac ctc atc cgc acg gat gaa aag ctt 192Gly Ser Leu Val Lys Lys Gly Asp Leu Ile Arg Thr Asp Glu Lys Leu 50 55 60gtc gaa atg cag aac ggc tgc att tgc tgt acg ctg aga gac gat ctg 240Val Glu Met Gln Asn Gly Cys Ile Cys Cys Thr Leu Arg Asp Asp Leu65 70 75 80ctc atc gaa gtc aaa aag ctg gcg gag aac ggc gga atc gat tat att 288Leu Ile Glu Val Lys Lys Leu Ala Glu Asn Gly Gly Ile Asp Tyr Ile 85 90 95ctc att gaa tct tcc ggt atc agc gag ccg att ccc gtc gcc caa acc 336Leu Ile Glu Ser Ser Gly Ile Ser Glu Pro Ile Pro Val Ala Gln Thr 100 105 110ttt acg tac aaa gat gag gag ctg aat att gat tta ggc aaa tat tgc 384Phe Thr Tyr Lys Asp Glu Glu Leu Asn Ile Asp Leu Gly Lys Tyr Cys 115 120 125agg ctt gat aca atg gtc acc gtc gtc gac ggc agc cgt ttt ttt gag 432Arg Leu Asp Thr Met Val Thr Val Val Asp Gly Ser Arg Phe Phe Glu 130 135 140gac ttc gct tcg ggc gag agc cta ttg gaa cgg aag cag gcg gca gat 480Asp Phe Ala Ser Gly Glu Ser Leu Leu Glu Arg Lys Gln Ala Ala Asp145 150 155 160gaa acg gac agc cgg gaa gtg gcc gac ctt tta att gag caa att gaa 528Glu Thr Asp Ser Arg Glu Val Ala Asp Leu Leu Ile Glu Gln Ile Glu 165 170 175ttt gcc gat gtc ctc att ttg aat aaa act gat tta ctc aat gaa gaa 576Phe Ala Asp Val Leu Ile Leu Asn Lys Thr Asp Leu Leu Asn Glu Glu 180 185 190gag cag gaa aag ctg ttc tgg tcc ctg aga aaa ctg aat ccg gac tgc 624Glu Gln Glu Lys Leu Phe Trp Ser Leu Arg Lys Leu Asn Pro Asp Cys 195 200 205aga atc att tct gcg gca ttc gca aaa gtg ccg ctt tcg gcc atc atg 672Arg Ile Ile Ser Ala Ala Phe Ala Lys Val Pro Leu Ser Ala Ile Met 210 215 220aat acg aaa tcg ttc gat ttt gaa aaa gca agt caa tca gcg gga tgg 720Asn Thr Lys Ser Phe Asp Phe Glu Lys Ala Ser Gln Ser Ala Gly Trp225 230 235 240atc aaa gag ctg aat gaa gag cac aca cct gaa aca gag gaa ttc gga 768Ile Lys Glu Leu Asn Glu Glu His Thr Pro Glu Thr Glu Glu Phe Gly 245 250 255atc tct tct ttc gtc tat caa aag caa agg cct ttc cat cct gaa aga 816Ile Ser Ser Phe Val Tyr Gln Lys Gln Arg Pro Phe His Pro Glu Arg 260 265 270ctg aag cgc tgg ctt gaa aac tgg ccg ccg caa att atc aga gca aaa 864Leu Lys Arg Trp Leu Glu Asn Trp Pro Pro Gln Ile Ile Arg Ala Lys 275 280 285ggc ttc ttc tgg ctt gct tct cga aat gac aca gca ggc ctt ctg tcc 912Gly Phe Phe Trp Leu Ala Ser Arg Asn Asp Thr Ala Gly Leu Leu Ser 290 295 300cag gca ggc tct tcc ctg atg atc caa agc gcg ggc ccc tgg att gca 960Gln Ala Gly Ser Ser Leu Met Ile Gln Ser Ala Gly Pro Trp Ile Ala305 310 315 320gcg tct acc gag gaa gaa caa aag ctc gcg ttt gaa gag gac cct aat 1008Ala Ser Thr Glu Glu Glu Gln Lys Leu Ala Phe Glu Glu Asp Pro Asn 325 330 335ttg aaa gcg gac tgg gat gaa aca tac gga gac cgg aaa acg gaa atc 1056Leu Lys Ala Asp Trp Asp Glu Thr Tyr Gly Asp Arg Lys Thr Glu Ile 340 345 350gtt ttt atc ggc att gac atg aat cag gaa gac att atc cgc tcc ctg 1104Val Phe Ile Gly Ile Asp Met Asn Gln Glu Asp Ile Ile Arg Ser Leu 355 360 365gat cag acc ctt ctt aca gat aaa gaa atg aat tca gac tgg tcg gcc 1152Asp Gln Thr Leu Leu Thr Asp Lys Glu Met Asn Ser Asp Trp Ser Ala 370 375 380tta ctt gat tca ctg cct gag tat att taa 1182Leu Leu Asp Ser Leu Pro Glu Tyr Ile385 39092393PRTBacillus licheniformis DSM 13 92Met Lys Glu Asn Leu Ile Pro Val Thr Val Leu Ser Gly Tyr Leu Gly1 5 10 15Ser Gly Lys Thr Thr Leu Leu Gln His Val Leu His Glu Ala His Gly 20 25 30Leu Lys Ile Ala Val Ile Val Asn Asp Met Ser Glu Ile Asn Ile Asp 35 40 45Gly Ser Leu Val Lys Lys Gly Asp Leu Ile Arg Thr Asp Glu Lys Leu 50 55 60Val Glu Met Gln Asn Gly Cys Ile Cys Cys Thr Leu Arg Asp Asp Leu65 70 75 80Leu Ile Glu Val Lys Lys Leu Ala Glu Asn Gly Gly Ile Asp Tyr Ile 85 90 95Leu Ile Glu Ser Ser Gly Ile Ser Glu Pro Ile Pro Val Ala Gln Thr 100 105 110Phe Thr Tyr Lys Asp Glu Glu Leu Asn Ile Asp Leu Gly Lys Tyr Cys 115 120 125Arg Leu Asp Thr Met Val Thr Val Val Asp Gly Ser Arg Phe Phe Glu 130 135 140Asp Phe Ala Ser Gly Glu Ser Leu Leu Glu Arg Lys Gln Ala Ala Asp145 150 155 160Glu Thr Asp Ser Arg Glu Val Ala Asp Leu Leu Ile Glu Gln Ile Glu 165 170 175Phe Ala Asp Val Leu Ile Leu Asn Lys Thr Asp Leu Leu Asn Glu Glu 180 185 190Glu Gln Glu Lys Leu Phe Trp Ser Leu Arg Lys Leu Asn Pro Asp Cys 195 200 205Arg Ile Ile Ser Ala Ala Phe Ala Lys Val Pro Leu Ser Ala Ile Met 210 215 220Asn Thr Lys Ser Phe Asp Phe Glu Lys Ala Ser Gln Ser Ala Gly Trp225 230 235 240Ile Lys Glu Leu Asn Glu Glu His Thr Pro Glu Thr Glu Glu Phe Gly 245 250 255Ile Ser Ser Phe Val Tyr Gln Lys Gln Arg Pro Phe His Pro Glu Arg 260 265 270Leu Lys Arg Trp Leu Glu Asn Trp Pro Pro Gln Ile Ile Arg Ala Lys 275 280 285Gly Phe Phe Trp Leu Ala Ser Arg Asn Asp Thr Ala Gly Leu Leu Ser 290 295 300Gln Ala Gly Ser Ser Leu Met Ile Gln Ser Ala Gly Pro Trp Ile Ala305 310 315 320Ala Ser Thr Glu Glu Glu Gln Lys Leu Ala Phe Glu Glu Asp Pro Asn 325 330 335Leu Lys Ala Asp Trp Asp Glu Thr Tyr Gly Asp Arg Lys Thr Glu Ile 340 345 350Val Phe Ile Gly Ile Asp Met Asn Gln Glu Asp Ile Ile Arg Ser Leu 355 360 365Asp Gln Thr Leu Leu Thr Asp Lys Glu Met Asn Ser Asp Trp Ser Ala 370 375 380Leu Leu Asp Ser Leu Pro Glu Tyr Ile385 39093390DNABacillus licheniformis DSM 13gene(1)..(390)putative protein - putative nitrogen regulatory protein P-II - BLi01176 93gtg aaa aaa atc gaa acg att ata cgc ccc gaa cag ttc ccc cgc ctc 48Val Lys Lys Ile Glu Thr Ile Ile Arg Pro Glu Gln Phe Pro Arg Leu1 5 10 15cgc aaa aag ctt gaa gag gtt ggc ata aac gga tta acc gtc tca gag 96Arg Lys Lys Leu Glu Glu Val Gly Ile Asn Gly Leu Thr Val Ser Glu 20 25 30gtt gcc ggc tgc ggc cag cag cgg ggg aag cag gct ctg ttt cgc ggc 144Val Ala Gly Cys Gly Gln Gln Arg Gly Lys Gln Ala Leu Phe Arg Gly 35 40 45acc tcc tac gaa atc aag ctg ctg cct aaa gtg aaa gtc gaa atg gtg 192Thr Ser Tyr Glu Ile Lys Leu Leu Pro Lys Val Lys Val Glu Met Val 50 55 60att gaa tcg gag agc gtt gat gaa att atc aaa tcg aaa gca cat gtt 240Ile Glu Ser Glu Ser Val Asp Glu Ile Ile Lys Ser Lys Ala His Val65 70 75 80cga caa ata cgg tcg gcg acg gaa aaa tct ttg tcc ttc caa ttg aaa 288Arg Gln Ile Arg Ser Ala Thr Glu Lys Ser Leu

Ser Phe Gln Leu Lys 85 90 95acg cga tcc gaa tcc gga cag ggg aat cag gaa agg aag cga ttg tct 336Thr Arg Ser Glu Ser Gly Gln Gly Asn Gln Glu Arg Lys Arg Leu Ser 100 105 110aaa acg acc gat tcc ctc ctg ctc tct ttt agt cga tac ttt aag aaa 384Lys Thr Thr Asp Ser Leu Leu Leu Ser Phe Ser Arg Tyr Phe Lys Lys 115 120 125ggg tga 390Gly 94129PRTBacillus licheniformis DSM 13misc_feature(1)..(1)Xaa can be Val or Met. 94Xaa Lys Lys Ile Glu Thr Ile Ile Arg Pro Glu Gln Phe Pro Arg Leu1 5 10 15Arg Lys Lys Leu Glu Glu Val Gly Ile Asn Gly Leu Thr Val Ser Glu 20 25 30Val Ala Gly Cys Gly Gln Gln Arg Gly Lys Gln Ala Leu Phe Arg Gly 35 40 45Thr Ser Tyr Glu Ile Lys Leu Leu Pro Lys Val Lys Val Glu Met Val 50 55 60Ile Glu Ser Glu Ser Val Asp Glu Ile Ile Lys Ser Lys Ala His Val65 70 75 80Arg Gln Ile Arg Ser Ala Thr Glu Lys Ser Leu Ser Phe Gln Leu Lys 85 90 95Thr Arg Ser Glu Ser Gly Gln Gly Asn Gln Glu Arg Lys Arg Leu Ser 100 105 110Lys Thr Thr Asp Ser Leu Leu Leu Ser Phe Ser Arg Tyr Phe Lys Lys 115 120 125Gly 95366DNABacillus licheniformis DSM 13gene(1)..(366)hypothetical protein - unknown function - BLi00719 95gtg aat caa ata cag ctg gaa gaa aag ttg tct att tta cta acg agt 48Val Asn Gln Ile Gln Leu Glu Glu Lys Leu Ser Ile Leu Leu Thr Ser1 5 10 15cga gtg att aca aaa aat gct ttc aac aca aca ttg aac aca ttt gtt 96Arg Val Ile Thr Lys Asn Ala Phe Asn Thr Thr Leu Asn Thr Phe Val 20 25 30ttc tta tca gaa aag ctg aaa aag gag gtg atc aaa gac tct gaa atg 144Phe Leu Ser Glu Lys Leu Lys Lys Glu Val Ile Lys Asp Ser Glu Met 35 40 45ttt tgg acg cac atg gcg atg gct tta act aga att aag gag gga gaa 192Phe Trp Thr His Met Ala Met Ala Leu Thr Arg Ile Lys Glu Gly Glu 50 55 60agc gaa gaa gtt cct gca aat gcc atc atc aat gaa atg tat gaa aca 240Ser Glu Glu Val Pro Ala Asn Ala Ile Ile Asn Glu Met Tyr Glu Thr65 70 75 80cca tat aaa agt gca ata gaa gaa gtg ata gac ttt ata cat tca ctg 288Pro Tyr Lys Ser Ala Ile Glu Glu Val Ile Asp Phe Ile His Ser Leu 85 90 95gca ggt cta gaa ctt cca gaa ggt gag ctc gtt tat ttt tat ctt cat 336Ala Gly Leu Glu Leu Pro Glu Gly Glu Leu Val Tyr Phe Tyr Leu His 100 105 110ctt cat aaa gtc ata gaa tct aat aaa tag 366Leu His Lys Val Ile Glu Ser Asn Lys 115 12096121PRTBacillus licheniformis DSM 13misc_feature(1)..(1)Xaa can be Val or Met. 96Xaa Asn Gln Ile Gln Leu Glu Glu Lys Leu Ser Ile Leu Leu Thr Ser1 5 10 15Arg Val Ile Thr Lys Asn Ala Phe Asn Thr Thr Leu Asn Thr Phe Val 20 25 30Phe Leu Ser Glu Lys Leu Lys Lys Glu Val Ile Lys Asp Ser Glu Met 35 40 45Phe Trp Thr His Met Ala Met Ala Leu Thr Arg Ile Lys Glu Gly Glu 50 55 60Ser Glu Glu Val Pro Ala Asn Ala Ile Ile Asn Glu Met Tyr Glu Thr65 70 75 80Pro Tyr Lys Ser Ala Ile Glu Glu Val Ile Asp Phe Ile His Ser Leu 85 90 95Ala Gly Leu Glu Leu Pro Glu Gly Glu Leu Val Tyr Phe Tyr Leu His 100 105 110Leu His Lys Val Ile Glu Ser Asn Lys 115 12097327DNABacillus licheniformis DSM 13gene(1)..(327)yvlA - unknown function - BLi03753 97atg acg aga aac aac gcg ctg att tct tca att tgt tat ttt agt gtt 48Met Thr Arg Asn Asn Ala Leu Ile Ser Ser Ile Cys Tyr Phe Ser Val1 5 10 15ttt ttt gcc ccg ctt att ttg cca atc gca gcg tat ttt ata aca gat 96Phe Phe Ala Pro Leu Ile Leu Pro Ile Ala Ala Tyr Phe Ile Thr Asp 20 25 30gat caa gaa acg aaa aag cat gcg atg cgt tcg ctt att tcg cat att 144Asp Gln Glu Thr Lys Lys His Ala Met Arg Ser Leu Ile Ser His Ile 35 40 45att ccg ttt ata agc gta gtc ata ttg gca gca ggc gcg ata agc ggt 192Ile Pro Phe Ile Ser Val Val Ile Leu Ala Ala Gly Ala Ile Ser Gly 50 55 60acg ctt atc ttt ggt gac ggc ggt tat ttg gcg gtc att ttc atg atc 240Thr Leu Ile Phe Gly Asp Gly Gly Tyr Leu Ala Val Ile Phe Met Ile65 70 75 80gga ggt ttc gtt ttt ttc ggg atc atc agt ttg att att gtg gtc tgg 288Gly Gly Phe Val Phe Phe Gly Ile Ile Ser Leu Ile Ile Val Val Trp 85 90 95aac gta gtt caa ggg att aaa atg ctg act cag gcc taa 327Asn Val Val Gln Gly Ile Lys Met Leu Thr Gln Ala 100 10598108PRTBacillus licheniformis DSM 13 98Met Thr Arg Asn Asn Ala Leu Ile Ser Ser Ile Cys Tyr Phe Ser Val1 5 10 15Phe Phe Ala Pro Leu Ile Leu Pro Ile Ala Ala Tyr Phe Ile Thr Asp 20 25 30Asp Gln Glu Thr Lys Lys His Ala Met Arg Ser Leu Ile Ser His Ile 35 40 45Ile Pro Phe Ile Ser Val Val Ile Leu Ala Ala Gly Ala Ile Ser Gly 50 55 60Thr Leu Ile Phe Gly Asp Gly Gly Tyr Leu Ala Val Ile Phe Met Ile65 70 75 80Gly Gly Phe Val Phe Phe Gly Ile Ile Ser Leu Ile Ile Val Val Trp 85 90 95Asn Val Val Gln Gly Ile Lys Met Leu Thr Gln Ala 100 10599333DNABacillus licheniformis DSM 13gene(1)..(333)tnrA - transcriptional pleiotropic regulator involved in global nitrogen regulation - BLi 01490 99atg aca gtc gaa gat ctt tct tat aaa gat aaa aaa gtg att tca att 48Met Thr Val Glu Asp Leu Ser Tyr Lys Asp Lys Lys Val Ile Ser Ile1 5 10 15ggc att gtc agc gag ctg acg ggt ctg tcg gtc agg cag atc cgc tat 96Gly Ile Val Ser Glu Leu Thr Gly Leu Ser Val Arg Gln Ile Arg Tyr 20 25 30tac gaa gaa cgc aag ctg atc tat ccg cag cgc tct aca aga ggc acg 144Tyr Glu Glu Arg Lys Leu Ile Tyr Pro Gln Arg Ser Thr Arg Gly Thr 35 40 45cgc aaa tat tcc ttt gct gac gtc gaa cgg ctg atg gaa atc gca aac 192Arg Lys Tyr Ser Phe Ala Asp Val Glu Arg Leu Met Glu Ile Ala Asn 50 55 60aag cgg gaa gac ggt gtt cag acc gca gaa atc ctc aaa gat atg cgc 240Lys Arg Glu Asp Gly Val Gln Thr Ala Glu Ile Leu Lys Asp Met Arg65 70 75 80aaa aaa gaa caa agg cta aaa aac gac cag cag ctc aga aag aaa atg 288Lys Lys Glu Gln Arg Leu Lys Asn Asp Gln Gln Leu Arg Lys Lys Met 85 90 95ctc gaa ggc cag ctt aac gct cat ttt aaa tac aaa aac cgt taa 333Leu Glu Gly Gln Leu Asn Ala His Phe Lys Tyr Lys Asn Arg 100 105 110100110PRTBacillus licheniformis DSM 13 100Met Thr Val Glu Asp Leu Ser Tyr Lys Asp Lys Lys Val Ile Ser Ile1 5 10 15Gly Ile Val Ser Glu Leu Thr Gly Leu Ser Val Arg Gln Ile Arg Tyr 20 25 30Tyr Glu Glu Arg Lys Leu Ile Tyr Pro Gln Arg Ser Thr Arg Gly Thr 35 40 45Arg Lys Tyr Ser Phe Ala Asp Val Glu Arg Leu Met Glu Ile Ala Asn 50 55 60Lys Arg Glu Asp Gly Val Gln Thr Ala Glu Ile Leu Lys Asp Met Arg65 70 75 80Lys Lys Glu Gln Arg Leu Lys Asn Asp Gln Gln Leu Arg Lys Lys Met 85 90 95Leu Glu Gly Gln Leu Asn Ala His Phe Lys Tyr Lys Asn Arg 100 105 110

Claims

1. A nucleic acid-binding chip, comprising probes for at least three of:kdgR, citA, gene coding for a putative protein (putative ABC transporter/amino acid permease) (homolog to SEQ ID NO. 91), htrA, ycnl, gene coding for a hypothetical protein of unknown function (homolog to SEQ ID NO. 41), gene coding for a hypothetical protein of unknown function (homolog to SEQ ID NO. 53), yppF, yqjN, ggt, alsR, glnA, nrgA, yciC, yvtA, nrgB, gene coding for a conserved hypothetical protein of unknown function (homolog to SEQ ID NO. 19), gene coding for a putative protein (ATP-binding protein of a putative ABC-transporter) (homolog to SEQ ID NO. 55), ycnJ, glnR, yvlA, yncE, yvlB, gene coding for a putative protein (putative hydrolase) (homolog to SEQ ID NO. 47), trpF, ydfS, trpD, gene coding for a putative protein (putative phage capsid protein) (homolog to SEQ ID NO. 21), ycnK, trpB, trpC, gene coding for a putative protein (putative transcription regulator) (homolog to SEQ ID NO. 17), gene coding for a putative protein (putative serine protease) (homolog to SEQ ID NO. 9), gene coding for a putative protein (putative glycosyl hydrolase/lysozyme) (homolog to SEQ ID NO. 85), gene coding for a putative protein (putative malate synthase, EC 4.1.3.2) (homolog to SEQ ID NO. 45), gene coding for a putative protein (putative Na(+)-bonded D-alanine-glycine permease) (homolog to SEQ ID NO. 49), gene coding for a hypothetical protein of unknown function (homolog to SEQ ID NO. 95), nasD, gene coding for a putative protein (putative ammonium transporter) (homolog to SEQ ID NO. 63), ycdH, nasC, gene coding for a hypothetical protein (close homolog to the aldehyde dehydrogenase DhaS) (homolog to SEQ ID NO. 15), nasB, trpE, pckA, gene coding for a putative protein (putative nitrogen regulation protein P-II) (homolog to SEQ ID NO. 93), nasF, yrkC, gene coding for a hypothetical protein of unknown function (homolog to SEQ ID NO. 81), or tnrA,wherein the total number of probes is not greater than 80.

2. The nucleic acid-binding chip according to claim 1, wherein the probes are selected from, in order of preference: kdgR, citA, gene coding for a putative protein (putative ABC transporter/amino acid permease) (homolog to SEQ ID NO. 91), htrA, ycnI, gene coding for a hypothetical protein of unknown function (homolog to SEQ ID NO. 41), gene coding for a hypothetical protein of unknown function (homolog to SEQ ID NO. 531 yppF, yqjN, ggt, alsR, glnA, nrgA, yciC, yvtA, nrgB, gene coding for a conserved hypothetical protein of unknown function (homolog to SEQ ID NO. 19), gene coding for a putative protein (ATP-binding protein of a putative ABC-transporter) (homolog to SEQ ID NO. 55), ycnJ, glnR, yvlA, yncE, yvlB, gene coding for a putative protein (putative hydrolase) (homolog to SEQ ID NO. 47), trpF, ydfS, trpD, gene coding for a putative protein (putative phage capsid protein) (homolog to SEQ ID NO. 21), ycnK, trpB, trpC, gene coding for a putative protein (putative transcription regulator) (homolog to SEQ ID NO. 17), gene coding for a putative protein (putative serine protease) (homolog to SEQ ID NO. 9), gene coding for a putative protein (putative glycosyl hydrolase/lysozyme) (homolog to SEQ ID NO. 85), gene coding for a putative protein (putative malate synthase, EC 4.1.3.2) (homolog to SEQ ID NO. 45), gene coding for a putative protein (putative Na(+)-bonded D-alanine-glycine permease) (homolog to SEQ ID NO. 49), gene coding for a hypothetical protein of unknown function (homolog to SEQ ID NO. 95), nasD, gene coding for a putative protein (putative ammonium transporter) (homolog to SEQ ID NO. 63), ycdH, nasC, gene coding for a hypothetical protein (close homolog to the aldehyde dehydrogenase DhaS) (homolog to SEQ ID NO. 15), nasB, trpE, pckA, gene coding for a putative protein (putative nitrogen regulation protein P-II) (homolog to SEQ ID NO. 93), nasF, yrkC, gene coding for a hypothetical protein of unknown function (homolog to SEQ ID NO. 81), and tnrA.

3. The nucleic acid-binding chip according to claim 1, wherein at least one of the probes is:gene coding for a conserved hypothetical protein of unknown function (homolog to SEQ ID NO. 19), gene coding for a putative protein (ATP-binding protein of a putative ABC-transporter) (homolog to SEQ ID NO. 55), ycnJ, glnR, yvlA, yncE, yvlB, gene coding for a putative protein (putative hydrolase) (homolog to SEQ ID NO. 47), trpF, ydfS, trpD, gene coding for a putative protein (putative phage capsid protein) (homolog to SEQ ID NO. 21), ycnK, trpB, trpC, gene coding for a putative protein (putative transcription regulator) (homolog to SEQ ID NO. 17), gene coding for a putative protein (putative serine protease) (homolog to SEQ ID NO. 9), gene coding for a putative protein (putative glycosyl hydrolase/lysozyme) (homolog to SEQ ID NO. 85), gene coding for a putative protein (putative malate synthase, EC 4.1.3.2) (homolog to SEQ ID NO. 45), gene coding for a putative protein (putative Na(+)-bonded D-alanine-glycine permease) (homolog to SEQ ID NO. 49), gene coding for a hypothetical protein of unknown function (homolog to SEQ ID NO. 95), nasD, gene coding for a putative protein (putative ammonium transporter) (homolog to SEQ ID NO. 63), ycdH, nasC, gene coding for a hypothetical protein (close homolog to the aldehyde dehydrogenase DhaS) (homolog to SEQ ID NO. 15), nasB, trpE, pckA, gene coding for a putative protein (putative nitrogen regulation protein P-II) (homolog to SEQ ID NO. 93), nasF, yrkC, gene coding for a hypothetical protein of unknown function (homolog to SEQ ID NO. 81), or tnrA.

4. The nucleic acid-binding chip according to claim 1 comprising at least 4 of the specified probes.

5. The nucleic acid-binding chip according to claim 1, wherein the total number of probes does not exceed 75, 70, 65, 60, 55, 50, 40, 30, 20 or 10.

6. (canceled)

7. (canceled)

8. The nucleic acid-binding chip according to claim 53, wherein the unicellular eukaryotes are protozoa or fungi.

9. The nucleic acid-binding chip according to claim 53, wherein the Gram-positive bacteria are Coryneform bacteria or a species of the genera Staphylococcus, Corynebacteria or Bacillus.

10. The nucleic acid-binding chip according to claim 53, wherein the Gram-negative bacteria are a species of the genera Escherichia or Klebsiella.

11. (canceled)

12. (canceled)

13. The nucleic acid-binding chip according to claim 1 further comprising at least one probe for detecting a gene encoding an amylase, cellulase, lipase, oxidoreductase, hemicellulase, or protease.

14. The nucleic acid-binding chip according to claim 1, wherein at least one of the probes is single-stranded.

15. The nucleic acid-binding chip according to claim 1, wherein at least one of the probes is DNA or a nucleic acid analog.

16. The nucleic acid-binding chip according to claim 1, wherein at least one of the probes comprises a region that is transcribed.

17. The nucleic acid-binding chip according to claim 1, wherein at least one of the probes is capable of binding to fragments of the gene of interest.

18. The nucleic acid-binding chip according to claim 1, wherein at least one of the probes is less than 200 nucleotides long.

19. The nucleic acid-binding chip according to claim 1, wherein an electric signal is triggered by the specific binding of mRNA to one or more probes.

20-27. (canceled)

28. A method for determining the physiological state of a unicellular eukaryote, a Gram positive bacteria, or a Gram negative bacteria under nitrogen deficiency conditions, comprising expression profiling the unicellular eukaryote, Gram positive bacteria, or Gram negative bacteria with the nucleic acid-binding chip of claim 1.

29. (canceled)

30. (canceled)

31. The method according to claim 28, wherein the unicellular eukaryote is a protozoa or fungi.

32. The method according to claim 28, wherein the Gram-positive bacteria are Coryneform bacteria or a species of the genera Staphylococcus, Corynebacteria or Bacillus.

33. The method according to claim 28, wherein the Gram-negative bacteria are a species of the genera Escherichia or Klebsiella.

34. The method according to claim 28, wherein the chip comprises probes derived from SEQ ID NOS: 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97 or 99.

35. The method according to claim 28, wherein the physiological state is determined at various times during nitrogen deficiency conditions.

36. The method according to claim 28, wherein the nitrogen deficiency condition occurs during fermentation.

37. The method according to claim 36, wherein the fermentation occurs during the production of a food or beverage product, nutritional supplement, or pharmaceutical by the unicellular eukaryote, Gram positive bacteria, or Gram negative bacteria.

38. The method according to claim 36, wherein the fermentation occurs during the expression of an α-amylase, protease, cellulase, lipase, oxidoreductase, peroxidase, laccase, oxidase or hemicellulase by the unicellular eukaryote, Gram positive bacteria, or Gram negative bacteria.

39-49. (canceled)

50. The nucleic acid binding chip according to claim 1, wherein at least one of the probes is trpC, gene coding for a putative protein (putative transcription regulator) (homolog to SEQ ID NO. 17), gene coding for a putative protein (putative serine protease) (homolog to SEQ ID NO. 9), gene coding for a putative protein (putative glycosyl hydrolase/lysozyme) (homolog to SEQ ID NO. 85), gene coding for a putative protein (putative malate synthase, EC 4.1.3.2) (homolog to SEQ ID NO. 45), gene coding for a putative protein (putative Na(+)-bonded D-alanine-glycine permease) (homolog to SEQ ID NO. 49), gene coding for a hypothetical protein of unknown function (homolog to SEQ ID NO. 95), nasD, gene coding for a putative protein (putative ammonium transporter) (homolog to SEQ ID NO. 63), ycdH, nasC, gene coding for a hypothetical protein (close homolog to the aldehyde dehydrogenase DhaS) (homolog to SEQ ID NO. 15), nasB, trpE, pckA, gene coding for a putative protein (putative nitrogen regulation protein P-II) (homolog to SEQ ID NO. 93), nasF, yrkC, gene coding for a hypothetical protein of unknown function (homolog to SEQ ID NO. 81), or tnrA.

51. The nucleic acid binding chip according to claim 1, wherein at least one of the probes is nasD, gene coding for a putative protein (putative ammonium transporter) (homolog to SEQ ID NO. 63), ycdH, nasC, gene coding for a hypothetical protein (close homolog to the aldehyde dehydrogenase DhaS) (homolog to SEQ ID NO. 15), nasB, trpE, pckA, gene coding for a putative protein (putative nitrogen regulation protein P-II) (homolog to SEQ ID NO. 93), nasF, yrkC, or gene coding for a hypothetical protein of unknown function (homolog to SEQ ID NO. 81).

52. The nucleic acid binding chip according to claim 1, wherein at least one of the probes is the gene coding for a putative protein (putative nitrogen regulation protein P-II) (homolog to SEQ ID NO. 93), nasF, yrkC, or gene coding for a hypothetical protein of unknown function (homolog to SEQ ID NO. 81).

53. The nucleic acid binding chip according to claim 1, wherein the probes correspond to a gene that exhibits at least an eight-fold change in expression in a unicellular eukaryote, Gram positive bacteria, or Gram negative bacteria in response to nitrogen deficiency conditions.

54. The nucleic acid binding chip according to claim 8, wherein the fungi are a species of Saccharomyces or Schizosaccharomyces.

55. The nucleic acid binding chip according to claim 9, wherein the Gram positive bacteria are Staphylococcus carnosus, Corynebacterium glutamicum, Bacillus subtilis, B. licheniformis, B. amyloliquefaciens, B. agaradherens, B. stearothermophilus, B. globigii B. lentus, or B. licheniformis.

56. The nucleic acid binding chip according to claim 10, wherein the Gram negative bacteria are derivatives of Escherichia coli K12, of Escherichia coli B, Klebsiella planticola, derivatives of Escherichia coli BL21 (DE3), E. coli RV308, E. coli DH5.alpha., E. coli JM109, E. coli XL-1 or Klebsiella planticola (Rf).

57. The nucleic acid binding chip according to claim 1, wherein at least one of the probes is less than 100 nucleotides long.

58. The nucleic acid binding chip according to claim 1, wherein at least one of the probes is 20 to 60 nucleotides long.

59. The nucleic acid binding chip according to claim 1, wherein at least one of the probes is 45 to 55 nucleotides long.

60. The method according to claim 31, wherein the fungi are a species of Saccharomyces or Schizosaccharomyces.

61. The method according to claim 32, wherein the Gram positive bacteria are Staphylococcus carnosus, Corynebacterium glutamicum, Bacillus subtilis, B. licheniformis, B. amyloliquefaciens, B. agaradherens, B. stearothermophilus, B. globigii B. lentus, or B. licheniformis.

62. The method according to claim 33, wherein the Gram negative bacteria are derivatives of Escherichia coli K12, of Escherichia coli B, Klebsiella planticola, derivatives of Escherichia coli BL21 (DE3), E. coli RV308, E. coli DH5.alpha., E. coli JM109, E. coli XL-1 or Klebsiella planticola (Rf).

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